Chib1和PAL5基因在AM真菌Glomus fasciculatus诱导大豆抗线虫病害防御反应中的作用

本研究系统分析了大豆（品种：‘鲁豆4’）接种AM真菌Glomus fasciculatum和胞囊线虫（SCN,Heterodera glycines）4号生理小种后各处理菌根和线虫侵染率、几丁质酶和苯丙氨酸解氨酶（PAL）活性及几丁质酶基因Chib1和苯丙氨酸解氨酶基因PAL5转录物的动态变化。结果表明,接种SCN对AM真菌的侵染率没有产生显著影响,但先接种AM真菌后接种SCN的大豆根内线虫侵染率明显低于只接种SCN的处理。另外,先接种AM真菌后接种SCN的大豆根内几丁质酶和PAL活性显著提高,活性高峰出现在接种线虫后的第3天。值得注意的是,先接种AM真菌后接种SCN的大豆根内两种基因Chib1和PAL5转录物高峰也出现在接种SCN后的第3天,即AM真菌侵染率快速上升而SCN侵染率快速下降时期。所以Chib1和PAL5基因的表达可能是AM真菌诱导的抗大豆胞囊线虫病害防御反应的一种表现。因此推测Chib1和PAL5直接参与了AM真菌诱导大豆抗胞囊线虫病害的防御反应。     

AM真菌和胞囊线虫对大豆根内酶活性的影响*

将‘鲁豆4号’大豆接种丛枝菌根(AM)真菌聚生球囊霉Glomus fasiculatum和大豆胞囊线虫(SCN)Heterodera glycines 4号生理小种后, 定期测定大豆根系中AM真菌及线虫侵染速率、过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)、β-1,3葡聚糖酶及几丁质酶活性的动态变化。结果表明, 接种AM真菌大豆根系中4种酶活性高于对照水平; 先接种AM真菌后接种SCN处理根系中POD、PAL及几丁质酶的活性高于只接种SCN的处理,并且酶活性峰值出现的时间均早于或相当于后者。另外,PAL及几丁质酶活性出现高峰时期也正是AM真菌侵染率迅速升高及线虫侵染速率快速下降期。因此,AM真菌先激活了大豆的防御反应,然后使其对SCN的侵染产生快速反应,PAL及几丁质酶在AM真菌诱导的抗、耐线虫病害机制中起重要作用。值得注意的是,先接种AM真菌后接种SCN处理大豆根系中,β-1,3葡聚糖酶活性低于只接种AM真菌的处理。作者认为本试验条件下,该酶在大豆抗SCN病害中的作用表现不明显。     

Effects on plant growth and root-knot nematode infection of an endophytic GFP transformant of the nematophagous fungus Pochonia chlamydosporia

Pochonia chlamydosporia (Pc123) is a fungal parasite of nematode eggs which can colonize endophytically barley and tomato roots. In this paper we use culturing as well as quantitative PCR (qPCR) methods and a stable GFP transformant (Pc123gfp) to analyze the endophytic behavior of the fungus in tomato roots. We found no differences between virulence/root colonization of Pc123 and Pc123gfp on root-knot nematode Meloidogyne javanica eggs and tomato seedlings respectively. Confocal microscopy of Pc123gfp infecting M. javanica eggs revealed details of the process such as penetration hyphae in the egg shell or appressoria and associated post infection hyphae previously unseen. Pc123gfp colonization of tomato roots was low close to the root cap, but increased with the distance to form a patchy hyphal network. Pc123gfp colonized epidermal and cortex tomato root cells and induced plant defenses (papillae). qPCR unlike culturing revealed reduction in fungus root colonization (total and endophytic) with plant development. Pc123gfp was found by qPCR less rhizosphere competent than Pc123. Endophytic colonization by Pc123gfp promoted growth of both roots and shoots of tomato plants vs. uninoculated (control) plants. Tomato roots endophytically colonized by Pc123gfp and inoculated with M. javanica juveniles developed galls and egg masses which were colonized by the fungus. Our results suggest that endophytic colonization of tomato roots by P. chlamydosporia may be relevant for promoting plant growth and perhaps affect managing of root-knot nematode infestations.     

丛枝菌根真菌摩西管柄囊霉侵染增强番茄对机械损伤的响应

植食性昆虫取食会给植物造成机械损伤并激活植物的防御反应,而与有益微生物共生是否可以增强植物对机械损伤的响应对植物抗虫有重要意义.本研究在番茄根系被丛枝菌根真菌摩西管柄囊霉侵染后,研究机械损伤对番茄防御反应的影响.结果表明: 预先接种菌根真菌的番茄叶片受到机械损伤处理(FD)后,叶片苯丙氨酸解氨酶(PAL)、超氧化物歧化酶(SOD)、过氧化物酶(POD)、多酚氧化酶(PPO)和过氧化氢酶(CAT)活性,以及叶片和根系苯丙氨酸解氨酶基因(PAL)和β-1,3-葡聚糖酶基因(PR2)的转录水平均显著高于只进行机械损伤的处理(D)、只接种摩西管柄囊霉的处理(F),以及既未接种菌根菌也未进行机械损伤的健康番茄植株(CK).虽然D和 F处理也可诱导部分酶活性及基因转录水平升高,但FD处理诱导的防御反应更迅速和强烈.表明丛枝菌根真菌侵染可以警备(prime)番茄对机械损伤做出更快速和强烈的响应.     

Evidence of differences between the communities of arbuscular mycorrhizal fungi colonizing galls and roots of Prunus persica infected by the root-knot nematode Meloidogyne incognita

Arbuscular mycorrhizal fungi (AMF) play important roles as plant protection agents, reducing or suppressing nematode colonization. However, it has never been investigated whether the galls produced in roots by nematode infection are colonized by AMF. This study tested whether galls produced by Meloidogyne incognita infection in Prunus persica roots are colonized by AMF. We also determined the changes in AMF composition and biodiversity mediated by infection with this root-knot nematode. DNA from galls and roots of plants infected by M. incognita and from roots of noninfected plants was extracted, amplified, cloned, and sequenced using AMF-specific primers. Phylogenetic analysis using the small-subunit (SSU) ribosomal DNA (rDNA) data set revealed 22 different AMF sequence types (17 Glomus sequence types, 3 Paraglomus sequence types, 1 Scutellospora sequence type, and 1 Acaulospora sequence type). The highest AMF diversity was found in uninfected roots, followed by infected roots and galls. This study indicates that the galls produced in P. persica roots due to infection with M. incognita were colonized extensively by a community of AMF, belonging to the families Paraglomeraceae and Glomeraceae, that was different from the community detected in roots. Although the function of the AMF in the galls is still unknown, we hypothesize that they act as protection agents against opportunistic pathogens.     

Arbuscular mycorrhizal fungi affect both penetration and further life stage development of root-knot nematodes in tomato

The root-knot nematode Meloidogyne incognita poses a worldwide threat to agriculture, with an increasing demand for alternative control options since most common nematicides are being withdrawn due to environmental concerns. The biocontrol potential of arbuscular mycorrhizal fungi (AMF) against plant-parasitic nematodes has been demonstrated, but the modes of action remain to be unraveled. In this study, M. incognita penetration of second-stage juveniles at 4, 8 and 12 days after inoculation was compared in tomato roots (Solanum lycopersicum cv. Marmande) pre-colonized or not by the AMF Glomus mosseae. Further life stage development of the juveniles was also observed in both control and mycorrhizal roots at 12 days, 3 weeks and 4 weeks after inoculation by means of acid fuchsin staining. Penetration was significantly lower in mycorrhizal roots, with a reduction up to 32%. Significantly lower numbers of third- and fourth-stage juveniles and females accumulated in mycorrhizal roots, at a slower rate than in control roots. The results show for the first time that G. mosseae continuously suppresses root-knot nematodes throughout their entire early infection phase of root penetration and subsequent life stage development.     

内生型解淀粉芽孢杆菌YTB1407对甘薯根系的促生作用及其调控机理

本实验室从西洋参根中分离筛选获得兼具生防和促生效果的内生型解淀粉芽孢杆菌YTB1407.为调查其应用潜力,以甘薯为材料,以无菌水灌根处理(CK)为对照,采用不同浓度YTB1407菌悬液灌根盆栽甘薯植株,通过对甘薯生长前期3个主要生育时间点YTB1407在植株内的内生定殖检测和根系表型效应比较,筛选菌悬液灌根的最适处理浓度.对根系内源生长素吲哚乙酸(IAA)、细胞分裂素玉米素核苷(ZR)、反式玉米素核糖核苷(t-ZR)的含量和吲哚乙酸氧化酶类中的吲哚乙酸氧化酶(IAAO)、过氧化物酶(POD)和多酚氧化酶(PPO)的活性进行分析.结果表明: YTB1407促进了甘薯生长前期根系统的定殖、不定根幼根的伸长生长及侧根的发生,提高了根系活力;在甘薯生长后期形成了更大的吸收根系生物量和更低的地上部和总根系生物量比.其中OD₆₀₀值为0.50的菌悬液处理(T_0.50)促生效应显著,在甘薯茎叶封垄期的单株块根鲜质量和有效薯块数量最高.YTB1407通过提高ZR、t-ZR含量,降低IAAO活性、提高PPO活性和IAA含量以及(t-ZR+ZR)/IAA,促进块根分化建成初期甘薯不定根幼根向块根的分化;通过降低IAAO、PPO活性和提高IAA含量,降低(t-ZR+ZR)/IAA,促进块根分化建成后期甘薯分化根向块根的建成.     

Trichoderma and chitin mixture based bioformulation for the management of head rot (Sclerotinia sclerotiorum (Lib.) deBary)–root-knot (Meloidogyne incognita Kofoid and White; Chitwood) complex diseases of cabbage

The effect of application of different biocontrol agents (Trichoderma spp (eight isolates), Pseudomonas fluorescens (four isolates) and Bacillus subtilis (two isolates)) was tested against head rot fungus, Sclerotinia sclerotiorum and root-knot nematode, Meloidogyne incognita complex diseases. In vitro studies showed that three Trichoderma isolates (Tvc1, Tvc2 and Thc) were effective in inhibition of mycelial growth of the fungus as well as egg hatching ability of the nematode. Application of talc based formulation of biocontrol agents (bioformulations) individually as well as in mixtures with or without chitin was tested against head rot–root-knot disease complex under greenhouse conditions. The combined application of bioformulation mixture (Tvc1, Tvc2 and Thc) along with chitin reduced the incidence of the complex diseases and induced significantly increased activities of phenylalanine ammonia-lyase (PAL), peroxidase (PO), polyphenol oxidase (PPO) and chitinase in cabbage. Activities of PAL and chitinase reached maximum levels within 10 – 20 d, while the activity of PAL continued to be maintained up to 40 d after the application of Tvc1 + Tvc2 + Thc + chitin. Isozyme analyses observed that unique PO (PO1, PO2 PO3 and PO4) and PPO (PPO1, PPO2 and PPO3) enzymes were induced after 10 d in cabbage plants treated with Tvc1 + Tvc2 + Thc + chitin upon challenge inoculation with head rot–root-knot pathogens. Similarly, the bioformulation mixture with chitin was successful at reducing the incidence of head rot–root-knot apart from enhancing the crop yield under field conditions. The mechanism associated with reduced incidence of head rot–root-knot in cabbage may be due to induction of defense proteins (PAL, PO, PPO and chitinase) in the crop.     

Interaction of Vesicular-Arbuscular Mycorrhizae and Cultivars of Alfalfa Susceptible and Resistant to Meloidogyne hapla

The interaction between vesicular-arbuscular mycorrhizal (VAM) fungi and the root-knot nematode (Meloidogyne hapla) was investigated using both nematode-susceptible (Grasslands Wairau) and nematode-resistant (Nevada Synthetic XX) cultivars of alfalfa (Medicago sativa) at four levels of applied phosphate. Mycorrhizal inoculation improved plant growth and reduced nematode numbers and adult development in roots in dually infected cultures of the susceptible cultivar. The tolerance of plants to nematode infection and development when preinfected with mycorrhizal fungi was no greater than when they were inoculated with nematodes and mycorrhizal fungi simultaneously. Growth of plants of the resistant cultivar was unaffected by nematode inoculation but was improved by mycorrhizal inoculation. Numbers of nematode juveniles were lower in the roots of the resistant than of the susceptible cultivar and were further reduced by mycorrhizal inoculation, although no adult nematodes developed in any resistant cultivar treatment. Inoculation of alfalfa with VAM fungi increased the tolerance and resistance of a cultivar susceptible to M. hapla and improved the resistance of a resistant cultivar.     

The effects of <Emphasis Type="Italic">β</Emphasis>-amino-butyric acid on resistance of cucumber against root-knot nematode, <Emphasis Type="Italic">Meloidogyne javanica</Emphasis>

In this study, the effects of β-amino-butyric acid (BABA) on root-knot nematode(Meloidogyne javanica) infection of cucumber and accumulation of total phenolic compounds, hydrogen peroxide and activity of some enzymes related to plant defense mechanisms, i.e., guaiacol peroxidase (GPOX), polyphenol oxidase (PPO), catalase (CAT) in cucumber roots infected with nematode were investigated. Results of this study show that treating the cucumber seedlings with the above elicitor significantly reduces the nematode infection level (the nematode galls, number of egg masses per plant and number of eggs per individual egg mass) compared to control. Additionally, treatment of cucumber roots by BABA and BABA + nematode, significantly increased peroxidase, polyphenol oxidase and catalase activities in root tissues, 1 day after nematode inoculation in comparison to nematode inoculated plants as control and sterile water-treated plants. Enzyme activities reached to a maximum level at 4, 4 and 3 days after nematode inoculation, respectively. Additionally, the amount of H₂O₂, a product of oxidative stress, was significantly increased in the BABA and BABA + nematode treatments in comparison to control. Such increases have occurred in two phases and maximum levels of it were observed at 5 days after inoculation. Inoculation of cucumber plants by BABA also significantly increased accumulation of total phenol in comparison to control and maximum level of it was observed at 7 days after nematode inoculation. The results suggest that the inhibitory effect of BABA on the root-knot nematode (M. javanica) may be related to its ability to enhance defense responses in the cucumber roots.     

Potential of Bacillus species against Meloidogyne javanica parasitizing eggplant (Solanum melongena L.) and induced biochemical changes

Aims

The biocontrol potential of three Bacillus species, namely Bacillus subtilis (BS), Bacillus firmus (BF), and Bacillus coagulans (BC) was tested against the root-knot nematode Meloidogyne javanica (Treub) Chitwood in eggplants (Solanum melongena L.). Plant growth and biochemical effects were also measured in these interactions.

Methods

Bacillus species were inoculated in soil around the seedlings of eggplants (Solanum melongena L.) with and without nematodes in a greenhouse experiment. Plant growth, biochemical changes, and nematode parasitism were observed at 15 and 45 days after inoculation (DAI).

Results

BC significantly enhanced plant growth, chlorophyll “b” and total chlorophyll contents, and polyphenol oxidase (PPO) activity in the leaves of eggplants, while BS showed greatest reduction in root-knot nematode parasitism. Non-infected and untreated control (C?) plants showed lesser chlorophyll “b,” carotenoids, soluble protein contents, and guaiacol peroxidase but higher catalase and PPO activities compared to infected and untreated controls (C+) at 15 and 45 DAI. Superoxide dismutase activity declined in most of the treated plants at 45 DAI following rise at 15 DAI. Ascorbate peroxidase activity increased at 45 DAI compared to 15 DAI in C? and C+ plants. PAL activity was greatly enhanced at 45 DAI in all treatments and controls over that at 15 DAI.

Conclusions

BC is a potentially plant growth-promoting bacteria although it was less effective against nematode infection compared to BS. Enzymes activities varied with infection and DAI. BC at 15 DAI in general increased the activity of most of the stress enzymes and thereby overcoming the effect of nematode parasitism.     

Influence of liming,inoculum level and inoculum placement on root colonization of subterranean clover

Arbuscular mycorrhizal (AM) fungi differ in their response to soil pH. Thus, change in soil pH may influence the relative abundance of mycorrhizal fungi inside roots. Root colonization by two AM fungi was studied in relation to addition of lime (CaCO3), quantity of inoculum and inoculum placement. Addition of CaCO3 to an acid soil decreased the colonization of roots by Acaulospora laevis but increased colonization by Glomus invermaium when both fungi were present. In acid soil (pH 4.7), almost all roots were colonized by A. laevis, while G. invermaium was dominant when soil pH was increased to pH 7.3. This occurred regardless of whether the inoculum was banded or mixed throughout the soil. There was no effect of CaCO3 on the relative abundance of fungi inside roots at intermediate rates of CaCO3 application (pH 5.3-6.3) when both fungi were inoculated together. In this experiment, both fungi colonized roots at all levels of CaCO3 when inoculated alone, except for A. laevis at the highest level of CaCO3. We conclude that soil pH affects the competitive ability of these two AM fungi during mycorrhiza formation primarily by affecting hyphae growth in soil and thus the relative abundance of hyphae at the root surface and subsequently inside the root.     

Auxins in the development of an arbuscular mycorrhizal symbiosis in maize

While the levels of free auxins in maize (Zea mays L.) roots during arbuscular mycorrhiza formation have been previously described in detail, conjugates of indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) with amino acids and sugars were neglected. In this study, we have therefore determined free, ester and amide bound auxins in roots of maize inoculated with Glomus intraradices during early stages of the colonization process. Ester conjugates of IAA and IBA were found only in low amounts and they did not increase in AM colonized roots. The Levels of IAA and IBA amide conjugates increased 20 and 30 days past inoculation (dpi). The formation of free and conjugated IBA but not IAA was systemically induced during AM colonization in leaves of maize plants. This implicated a role for auxin conjugate synthesis and hydrolysis during AM. We have therefore investigated the in vivo metabolism of 3H-labeled IBA by TLC but only slight differences between control and AM-inoculated roots were observed. The activity of auxin conjugate hydrolase activity measured with three different putative substrates showed a decrease in infected roots compared to controls. The fluorinated IBA analog TFIBA inhibited IBA formation in leaves after application to the root system, but was not transported from roots to shoots. AM hyphae were also not able to transport TFIBA. Our results indicate complex control mechanisms to regulate the levels of free and conjugated auxins, which are locally and systemically induced during early stages of the formation of an arbuscular mycorrhizal symbiosis.     

Interactions between the root-knot nematode Meloidogyne incognita and Glomus mosseae in banana

The effects of the interaction between the arbuscular mycorrhizal fungus Glomus mosseae and the root-knot nematode Meloidogyne incognita on growth and nutrition of micropropagated ;Grand Naine banana (Musa AAA) cultivar was studied under greenhouse conditions. Inoculation with two G. mosseae isolates significantly increased growth of plants in relation to non-mycorrhizal plants. Response to mycorrhizae was as effective as with an optimum P fertilization in promoting plant development for most growth parameters. Meloidogyne incognita had no apparent effect on the percentage of root colonization in mycorrhizal plants. In contrast, G. mosseae suppressed root galling and nematode buildup in the roots. The percentage of mycorrhizal colonization was high (over 80%) in low P fertilized plants, but optimum P rates for bananas (four times higher than low P) significantly reduced mycorrhizal colonization. Most elements were within sufficiency levels for banana with exception of N which was low for all treatments. Mycorrhizal plants fertilized with a low P rate showed higher N, P, K, Ca, and Mg contents as compared to non-mycorrhizal plants low in P with or without the nematode. Inoculation with G. mosseae favours growth of banana plants by enhancing plant nutrition and by suppressing nematode reproduction and galling during the early stages of plant development.     

Effectiveness of autochthonous bacterium and mycorrhizal fungus on Trifolium growth, symbiotic development and soil enzymatic activities in Zn contaminated soil

AIMS: This study investigates how autochthonous micro-organisms [bacterium and/or arbuscular mycorrhizal (AM) fungi] affected plant tolerance to Zn contamination. METHODS AND RESULTS: Zinc-adapted and -nonadapted Glomus mosseae strains protected the host plant against the detrimental effect of Zn (600 microg g(-1)). Zn-adapted bacteria increased root growth and N, P nutrition in plants colonized by adapted G. mosseae and decreased the specific absorption rate (SAR) of Cd, Cu, Mo or Fe in plants colonized by Zn-nonadapted G. mosseae. Symbiotic structures (nodule number and extraradical mycelium) were best developed in plants colonized by those Zn-adapted isolates that were the most effective in increasing plant Zn tolerance. The bacterium also increased the quantity and quality (metabolic characteristics) of mycorrhizal colonization, with the highest improvement for arbuscular vitality and activity. Inocula also enhanced soil enzymatic activities (dehydrogenase, beta-glucosidase and phosphatase) and indol acetic acid (IAA) accumulation, particularly in the rhizosphere of plants inoculated with Zn-adapted isolates. CONCLUSIONS: Glomus mosseae strains have a different inherent potential for improving plant growth and nutrition in Zn-contaminated soil. The bacterium increased the potential of mycorrhizal mycelium as inoculum. SIGNIFICANCE AND IMPACT OF THE STUDY: Mycorrhizal performance, particularly that of the autochthonous strain, was increased by the bacterium and both contributed to better plant growth and establishment in Zn-contaminated soils.     

Rhizobial Nodulation Factors Stimulate Mycorrhizal Colonization of Nodulating and Nonnodulating Soybeans

Legumes form tripartite symbiotic associations with noduleinducing rhizobia and vesicular-arbuscular mycorrhizal fungi. Co-inoculation of soybean (Glycine max [L.] Merr.) roots with Bradyrhizobium japonicum 61-A-101 considerably enhanced colonization by the mycorrhizal fungus Glomus mosseae. A similar stimulatory effect on mycorrhizal colonization was also observed in nonnodulating soybean mutants when inoculated with Bradyrhizobium japonicum and in wild-type soybean plants when inoculated with ineffective rhizobial strains, indicating that a functional rhizobial symbiosis is not necessary for enhanced mycorrhiza formation. Inoculation with the mutant Rhizobium sp. NGR[delta]nodABC, unable to produce nodulation (Nod) factors, did not show any effect on mycorrhiza. Highly purified Nod factors also increased the degree of mycorrhizal colonization. Nod factors from Rhizobium sp. NGR234 differed in their potential to promote fungal colonization. The acetylated factor NodNGR-V (MeFuc, Ac), added at concentrations as low as 10-9 M, was active, whereas the sulfated factor, NodNGR-V (MeFuc, S), was inactive. Several soybean flavonoids known to accumulate in response to the acetylated Nod factor showed a similar promoting effect on mycorrhiza. These results suggest that plant flavonoids mediate the Nod factor-induced stimulation of mycorrhizal colonization in soybean roots.     

Phosphorus and intraspecific density alter plant responses to arbuscular mycorrhizas

We investigated how phosphorus availability, intraspecific density, and their interaction affect plant responses to arbuscular mycorrhizas. Four facultatively mycotrophic species: chile, cilantro, tomato, and corn were examined separately in pot experiments that employed a tropical phosphorus-immobilizing soil. Each experiment comprised nine soluble phosphorus additions, two levels of intraspecific plant density, and inoculation with arbuscular mycorrhizal fungi or not. High phosphorus signi- ficantly diminished mycorrhizal colonization of corn, cilantro, and tomato, but not chile, which was highly variably colonized. Corn roots were colonized by other root-inhabiting fungi, and mycorrhizas significantly reduced colonization by these potential root parasites. High phosphorus significantly increased relative growth rates (RGR) of all species, and high density significantly reduced RGR of cilantro, tomato, and corn. Chile showed little growth at any but the highest phosphorus additions, and consequently had no RGR response to density or mycorrhizas. Mycorrhiza inoculation caused transient depression of corn growth during the first month, but mycorrhizas increased corn RGR during the second month of growth. Both RGR and dry weights at harvest, cilantro, tomato, and corn benefited from mycorrhizas at low phosphorus availability, but this benefit diminished or changed to disadvantage as phosphorus availability increased. At low phosphorus availability, high density increased the dry weight of mycorrhizal cilantro and thereby amplified the benefit of mycorrhizas. At high phosphorus availability, increased density diminished the effects of mycorrhizas on dry weight, reducing mycorrhiza benefit to tomato and chile and reducing mycorrhiza detriment to cilantro. This study demonstrates that for three of the four plant species examined, phosphorus availability, intraspecific density, and their interaction significantly modify plant responses to arbuscular mycorrhizas.     

Nitrogen assimilation in Lolium perenne colonized by the arbuscular mycorrhizal fungus Glomus fasciculatum

To investigate nitrogen assimilation in Lolium perenne L. colonized by the arbuscular mycorrhizal (AM) fungus Glomus fasciculatum (Thax. sensu Gerd.), nitrate uptake, key enzyme activities, and ¹⁵N incorporation into free amino acids were measured. After a 4-h labelling period with [¹⁵N]nitrate, ¹⁵N content was higher in roots and shoots of AM-plants than in those of control plants. Glutamine synthetase (GS) and nitrate reductase (NR) activities were increased in shoots of AM-plants, but not in roots. More label was incorporated into amino acids in shoots of AM plants. Glutamine, glutamate, alanine and γ-aminobutyric acid were the major sinks for ¹⁵N in roots and shoots of control and AM plants. Interactions between mycorrhizal colonization, phosphate and nitrate nutrition and NR activity were investigated in plants which received different amounts of phosphate or nitrate. In shoots of control plants, NR activity was not stimulated by high levels of phosphate nutrition but was stimulated by high levels of nitrate. At 4 m M nitrate in the nutrient solution, NR activity was similar in control and AM plants. We concluded that mycorrhizal effects on nitrate assimilation are not mediated via improved phosphate nutrition, but could be due to improved nitrogen uptake and translocation.     

The mobilisation and fate of soil and rock phosphate in the rhizosphere of ectomycorrhizal Pinus radiata seedlings in an Allophanic soil

A pot experiment was conducted to investigate the uptake of Zn from experimentally contaminated calcareous soil of low nutrient status by maize inoculated with the arbuscular mycorrhizal (AM) fungus Glomus caledonium. EDTA was applied to the soil to mobilize Zn and thus maximize plant Zn uptake. The highest plant dry matter (DM) yields were obtained with a moderate Zn addition level of 300 mg kg^?1. Plant growth was enhanced by mycorrhizal colonization when no Zn was added and under the highest Zn addition level of 600 mg kg^?1, while application of EDTA to the soil generally inhibited plant growth. EDTA application also increased plant Zn concentration, and Zn accumulation in the roots increased with increasing EDTA addition level. The effects of inoculation with G. caledonium on plant Zn uptake varied with Zn addition level. When no Zn was added, Zn translocation from roots to shoots was enhanced by mycorrhizal colonization. In contrast, when Zn was added to the soil, mycorrhizal colonization resulted in lower shoot Zn concentrations in mycorrhizal plants. The P nutrition of the maize was greatly affected by AM inoculation, with mycorrhizal plants showing higher P concentrations and P uptake. The results indicate that application of EDTA mobilized soil Zn, leading to increased Zn accumulation by the roots and subsequent plant toxicity and growth inhibition. Mycorrhizal colonization alleviated both Zn deficiency and Zn contamination, and also increased host plant growth by influencing mineral nutrition. However, neither EDTA application nor arbuscular mycorrhiza stimulated Zn translocation from roots to shoots or metal phytoextraction under the experimental conditions. The results are discussed in relation to the environmental risk associated with chelate-enhanced phytoextraction and the potential role of arbuscular mycorrhiza in soil remediation.