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.     

Network establishment of arbuscular mycorrhizal hyphae in the rhizospheres between citrus rootstocks and <Emphasis Type="Italic">Paspalum notatum</Emphasis> or <Emphasis Type="Italic">Vulpia myuros</Emphasis> grown in sand substrate

A greenhouse experiment was conducted to examine the favorable effects of sod culture system with bahiagrass (Paspalum notatum Flügge.) and Vulpia myuros (L.) C. C. Gmel. intercropped with citrus trees on the establishment of the network of arbuscular mycorrhizal (AM) fungus hyphae in their rhizospheres. Special acrylic root boxes with three compartments were used for the experiment. Four types of citrus rootstock seedlings, trifoliate orange (Poncirus trifoliata Raf.), sour orange (Citrus aurantium L.), rough lemon (Citrus jambhiri Lush.), and Citrus natsudaidai Hayata, were separately transplanted into one outer compartment in each box, and the seedlings of bahiagrass and V.myuros were separately transplanted into the other outer compartment. An AM fungus, Gigaspora margarita Becker and Hall, was inoculated in the center compartment of each box. Some boxes with both outer compartments without plants and with some plants in only one outer compartment were also prepared. The box with bare × bare had very low density of AM hyphae. There were a few hyphae in bare compartments in the boxes of trifoliate orange × bare, sour orange × bare, rough lemon × bare, and C. natsudaidai × bare. The density of hyphae in the compartments with citrus seedlings and grasses, however, was significantly higher than in every bare compartment, and the hyphae in the compartments with plants penetrated deeply into the sand. In particular, the density in the compartments of citrus seedlings increased when bahiagrass or V. myuros was transplanted as a neighboring plant. The percentage of AM fungus colonization in every plant root was high. New spore formation was observed in compartments with plants, whereas there were few spores in every bare compartment. In particular, the spore formation in bahiagrass compartments was superior to that in other compartments with plants. Our results suggest that the network system by AM hyphae is easily discernible in the rhizospheres between citrus rootstocks and bahiagrass or V. myuros, but bare ground severely inhibits the formation and development of AM hyphal network and reduces the number of AM spores in the soil.     

Influence of arbuscular mycorrhizal fungi and salinity on growth,biomass, and mineral nutrition of<Emphasis Type="Italic"> Acacia auriculiformis</Emphasis>

The effect of salinity on the efficacy of two arbuscular mycorrhizal fungi, Glomus fasciculatum and G. macrocarpum, alone and in combination was investigated on growth, development and nutrition of Acacia auriculiformis. Plants were grown under different salinity levels imposed by 0.3, 0.5 and 1.0 S m^-1 solutions of 1 M NaCl. Both mycorrhizal fungi protected the host plant against the detrimental effect of salinity. The extent of AM response on growth as well as root colonization varied with fungal species, and with the level of salinity. Maximum root colonization and spore production was observed with combined inoculation, which resulted in greater plant growth at all salinity levels. AM fungal inoculated plants showed significantly higher root and shoot weights. Greater nutrient acquisition, changes in root morphology, and electrical conductivity of soil in response to AM colonization was observed, and may be possible mechanisms to protect plants from salt stress.     

Interactions between arbuscular mycorrhizal fungi and phosphate-solubilizing fungus (<Emphasis Type="Italic">Mortierella</Emphasis> sp.) and their effects on <Emphasis Type="Italic">Kostelelzkya virginica</Emphasis> growth and enzyme activities of rhizosphere and bulk soils at different salinities

This study investigated the interactions between two arbuscular mycorrhizal fungi (AMF) (Glomus aggregatum and Glomus mosseae) and a P-solubilizing fungus (Mortierella sp.), with respect to their effects on growth of Kostelelzkya virginica and urease, invertase, neutral phosphatase, alkaline phosphatase, and catalase activities of rhizosphere and bulk soils at different salinity levels (i.e., 0, 100, 200, and 300 mM NaCl). Percentage of AMF colonization, Mortierella sp. populations, pH, electrical conductivity, and available P concentration in soil were also determined. Combined inoculation of AMF and Mortierella sp. increased the percentage of AMF colonization and Mortierella sp. populations under salt stress (i.e., 100, 200, and 300 mM NaCl). The dual inoculation of Mortierella sp. with AMF (G. aggregatum or G. mosseae) had significant effects on shoot and root dry weights and available P concentrations, pH values, and electrical conductivities of rhizosphere and bulk soils under salt stress. The inoculation of Mortierella sp. significantly enhanced the positive effects of AMF on some enzyme activities (i.e., neutral phosphatase, alkaline phosphatase, and catalase in bulk soil; neutral phosphatase and urease in rhizosphere soil); on the contrary, it produced negative effects on urease activities in bulk soil and invertase activities in bulk and rhizosphere soils. The results indicated that the most effective co-inoculation was the dual inoculation with Mortierella sp. and G. mosseae, which may help in alleviating the deleterious effects of salt on plants growth and soil enzyme activities.     

Production and cytogenetics of trigeneric hybrid involving <Emphasis Type="Italic">Triticum</Emphasis>, <Emphasis Type="Italic">Psathyrostachys</Emphasis> and <Emphasis Type="Italic">Secale</Emphasis>

Trigeneric hybrids may help establish evolutionary relationships among different genomes present in the same cellular-genetic background, and also offers the possibility to transfer different alien characters into cultivated wheat. In this study, a new trigeneric hybrid involving species from the Triticum, Psathyrostachys and Secale was synthesized by crossing wheat-P. huashanica amphiploid (PHW-SA) with wheat-S. cereale amphiploid (Zhongsi 828). The crossability of F₁ hybrid was high with 35.13%, and the fertility was 41.95%. The morphological characteristics of F₁ plants resembled the parent Zhongsi 828. The trigeneric hybrids pollen mother cells (PMCs) regularly revealed averagely 19.88 univalents, 9.63 ring bivalents, 3.97 rod bivalents, 0.60 trivalents and 0.03 tetravalents per cell. Multivalents consisted of trivalents and tetravalents can be observed in 52.7% of cells. A variation of abnormal lagging chromosome, micronuclei and chromosome bridge were formed at anaphase I and telophase II. The mean chromosomes number of F₂ progenies was 2n = 46.13, and the distribution range was 42–53. GISH results revealed that most F₂ plants had 6–12 S. cereale chromosomes, and only 0–2 P. huashanica chromosomes were detected. The results indicated that S. cereale chromosomes can be preferentially transmitted in the F₂ progenies of trigeneric hybrid than P. huashanica chromosomes. A survey of disease resistances revealed that the stripe rust resistance from the PHW-SA were completely expressed in the F₁ and some F₂ plants. The trigeneric hybrid could be a useful bridge for the transference of P. huashanica and S. cereale chromatins to common wheat.     

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 salinity and flooding on the infectivity of salt marsh arbuscular mycorrhizal fungi in<Emphasis Type="Italic"> Aster tripolium</Emphasis> L.

Salt marshes are characterized by the occurrence of combined salinity and flooding stresses. The individual and combined effects of salinity and flooding on the establishment and activity of arbuscular mycorrhizal (AM) colonization in the salt marsh halophyte Aster tripolium L. by indigenous salt marsh AM fungi were evaluated. A. tripolium plants were cultivated in a mixture of sand and salt marsh soil under different salinity concentrations (5%, 50% or 100% artificial seawater) and water regimes (non-flooding, tidal flooding and continuous flooding). Plants were harvested after 3 and 8 weeks and their growth was negatively influenced by increased salinity and water level. Increased salinity level affected the establishment of AM colonization, AM fungal growth and activity (measured as succinate dehydrogenase activity) within roots, and extraradical mycelium growth. The influence of flooding on the establishment of colonization and on intra- and extraradical AM fungal growth was dependent on the water regime. Continuous flooding reduced colonization and AM fungal growth, whereas tidal flooding did not affect these parameters unless combined with intermediate salinity level (50% seawater) at the end of the experiment. The water regime did not influence AM active colonization. The ratio of root to soil AM fungal growth increased as the water level increased. The results of this study demonstrate that the establishment and activity of AM colonization in A. tripolium is more influenced by salinity than by flooding, and suggests that the functionality of salt marsh AM fungi is not affected by flooding.     

Physiological Effects and Fluorescence Labeling of Magnetic Iron Oxide Nanoparticles on Citrus (<Emphasis Type="Italic">Citrus reticulata</Emphasis>) Seedlings

Nanoparticles (NPs) have been reported to cause physiological effects on plant cells and tissue. This study traced the uptake and distribution of magnetic iron oxide nanoparticles (γ-Fe₂O₃ NPs) in citrus (Citrus reticulata) plants under hydroponic condition by fluorescent dye labeled γ-Fe₂O₃ NPs, and described a detailed evidence of physiological effects of 0–100 mg/L γ-Fe₂O₃ NPs on citrus plants by measuring the physiological parameters such as content of chlorophyll, malondialdehyde (MDA), soluble sugar, soluble protein, activity of antioxidant enzyme, and ferric reductase after 21 days exposure. Fluorescence images of citrus stem and root showed that citrus roots could absorb γ-Fe₂O₃ NPs but no translocation from roots to shoots was observed, since NPs aggregated or even clogged the vascular system. Physiological results showed that 20 mg/L γ-Fe₂O₃ NPs could significantly enhance chlorophyll content by 126.4%, while 50 and 100 mg/L of γ-Fe₂O₃ NPs decreased chlorophyll content by 27.8 and 35.4%, respectively. MDA contents in citrus leaves under 20–100 mg/L γ-Fe₂O₃ NPs exposure were increased by 37.8, 107.2, and 61.5%, respectively, while that in roots were decreased by 27.0,11.9, and 7.4%, respectively, with elevated SOD and CAT activity, suggesting that oxidative stress occurred in citrus leaves, but oxidative stress in roots was eliminated by antioxidant defense. It is noteworthy that although Fe(II)-EDTA treatment had a high level of chlorophyll content, it induced strong oxidative stress in citrus plants as well. Collectively, the various physiological responses of citrus plants to γ-Fe₂O₃ NPs exposure were closely correlated with the concentrations of NPs. γ-Fe₂O₃ NPs at proper concentrations, such as 20 mg/L, have the potential to ameliorate chlorosis of plants and be effective nanofertilizers for increasing agronomic productivity.     

Arbuscular mycorrhizal fungi optimize the acquisition and translocation of Cd and P by cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis> L.) plant cultivated on a Cd-contaminated soil

Purpose

Fruiting vegetables are generally considered to be safer than other vegetables for planting on cadmium (Cd)-contaminated farms. However, the risk of transferring Cd that has accumulated in the stems and leaves of fruiting vegetables is a major issue encountered with the usage of such non-edible parts. The objective of this study was to resolve the contribution of arbuscular mycorrhizal (AM) fungi to the production of low-Cd fruiting vegetables (focusing on the non-edible parts) on Cd-contaminated fields.

Materials and methods

An 8-week pot experiment was conducted to investigate the acquisition and translocation of Cd by cucumber (Cucumis sativus L.) plants on an unsterilized Cd-contaminated (1.6 mg kg^?1) soil in response to inoculation with the AM fungus, Funneliformis caledonium (Fc) or Glomus versiforme (Gv). Mycorrhizal colonization rates of cucumber roots were assessed. Dry biomass and Cd and phosphorus (P) concentrations in the cucumber shoots and roots were all measured. Soil pH, EC, total Cd, phytoavailable (DTPA-extractable) Cd, available P, and acid phosphatase activity were also tested.

Results and discussion

Both Fc and Gv significantly increased (P?<?0.05) root mycorrhizal colonization rates and P acquisition efficiencies, and thus the total P acquisition and biomass of cucumber plants, whereas only Fc significantly increased (P?<?0.05) soil acid phosphatase activity and the available P concentration. Both Fc and Gv significantly increased (P?<?0.05) root to shoot P translocation factors, inducing significantly higher (P?<?0.05) shoot P concentrations and shoot/root biomass ratios. In contrast, both Fc and Gv significantly decreased (P?<?0.05) root and shoot Cd concentrations, resulting in significantly increased (P?<?0.05) P/Cd concentration ratios, whereas only Gv significantly decreased (P?<?0.05) the root Cd acquisition efficiency and increased (P?<?0.05) the root to shoot Cd translocation factor. Additionally, AM fungi also tended to decrease soil total and phytoavailable Cd concentrations by elevating plant total Cd acquisition and soil pH, respectively.

Conclusions

Inoculation with AM fungi increased the P acquisition and biomass of cucumber plants, but decreased plant Cd concentrations by reducing the root Cd acquisition efficiency, and resulted in a tendency toward decreases in soil phytoavailable and total Cd concentrations via increases in soil pH and total Cd acquisition by cucumber plants, respectively. These results demonstrate the potential application of AM fungi for the production of fruiting vegetables with non-edible parts that contain low Cd levels on Cd-contaminated soils.

     

Mitigation of salt stress in wheat seedlings by a <Emphasis Type="Italic">gfp</Emphasis>-tagged <Emphasis Type="Italic">Azospirillum lipoferum</Emphasis>

Root colonization and mitigation of NaCl stress on wheat seedlings were studied by inoculating seeds with Azospirillum lipoferum JA4ngfp15 tagged with the green fluorescent protein gene (gfp). Colonization of wheat roots under 80 and 160 mM NaCl stress was similar to root colonization with this bacterial species under non-saline conditions, that is, single cells and small aggregates were mainly located in the root hair zone. These salt concentrations had significant inhibitory effects on development of seedlings, but not on growth in culture of gfp-A. lipoferum JA4ngfp15. Reduced plant growth (height and dry weight of leaves and roots) under continuous irrigation with 160 mM NaCl was ameliorated by bacterial inoculation with gfp-A. lipoferum JA4ngfp15. Inoculation of plants subjected to continuous irrigation with 80 mM NaCl or to a single application of either NaCl concentration (80 or 160 mM NaCl) did not mitigate salt stress. This study indicates that, under high NaCl concentration, inoculation with modified A. lipoferum reduced the deleterious effects of NaCl; colonization patterns on roots were unaffected and the genetic marker did not induce undesirable effects on the interaction between the bacterium and the plants.     

Characterization of fertile amphidiploid between <Emphasis Type="Italic">Raphanus sativus</Emphasis> and <Emphasis Type="Italic">Brassica alboglabra</Emphasis> and the crossability with <Emphasis Type="Italic">Brassica</Emphasis> species

A fertile amphidiploid × Brassicoraphanus (RRCC, 2n = 36) between Raphanus sativus cv. HQ-04 (2n = 18, RR) and Brassica alboglabra Bailey (2n = 18, CC) was synthesized and successive selections for seed fertility were made from F₄ to F₁₀. F₁₀ plants exhibited good fertility with 14.9 seeds per siliqua and 32.3 g seeds per plant. Cytological observation revealed that frequent secondary pairing occurred among 3 chromosome pairs in pollen mother cells of plants (F₄) with lower fertility, but not of plants with high fertility (F₁₀). GISH analysis indicated that these F₁₀ plants included the expected 18 chromosomes from R. sativus and B. alboglabra, respectively, but they lost approximately 27.6% R. sativus and 35.6% B. alboglabra AFLP (amplified fragment length polymorphism) bands. The crossability of the Raphanobrassica with R. sativus and 5 Brassica species (13 cultivars) were investigated. Seeds or F₁ seedlings were easy to be produced from crosses × Brassicoraphanus × R. sativus, and B. napus, B. juncea and B. carinata × Brassicoraphanus. Fewer seeds or seedlings were obtained from crosses × Brassicoraphanus × B. napus, B. juncea and B. carinata. However, few seeds were harvested in the reciprocals of × Brassicoraphanus with B. rapa and B. oleracea. The possible cause of fertility improvements and the potential of the present × Brassicoraphanus for breeding were discussed.     

Influence of arbuscular mycorrhizal fungi,inoculum level and phosphorus placement on growth and phosphorus uptake of <Emphasis Type="Italic">Phyllanthus calycinus</Emphasis> under jarrah forest soil

Phosphorus uptake and biomass production may vary between arbuscular mycorrhizas (AM) species of contrasting life cycles and their modes of interaction with host plants. This research investigated differences among three AM fungi from different genera isolated from the jarrah forest with respect to their colonisation, biomass production and P uptake over time. The understorey plant Phyllanthus calycinus, has been shown to be extensively mycorrhizal. We examined differences in the capacity of fungi associated with this plant to access a P point source from increasing distances from the root. The methodology simulated “pockets” of P in forest soil. Phosphorus sources in root-exclusion mesh bags were inserted 2, 4 and 6 cm from plant roots restrained in separate mesh bags. Acaulospora laevis colonised plants to the greatest extent, producing higher plant biomass and P uptake compared to the other two fungi. Plant biomass and P uptake were enhanced where P sources were closest to the root, as expected, but there were differences among fungi in response to level of inoculum used for each fungus. The capacity of individual AM fungi to access a point source of P at different distances varied in a highly mycorrhizal native forest understorey plant in parallel with plant growth and P uptake.     

<Emphasis Type="Italic">Achromobacter insolitus</Emphasis> and <Emphasis Type="Italic">Zoogloea ramigera</Emphasis> associated with wheat plants (<Emphasis Type="Italic">Triticum aestivum</Emphasis>)

This study reports for the first time the presence of diazotrophic bacteria belonging to the genera Achromobacter and Zoogloea associated with wheat plants. These bacterial strains were identified by the analysis of 16S rDNA sequences. The bacterium IAC-AT-8 was identified as Azospirillum brasiliense, whereas isolates IAC-HT-11 and IAC-HT-12 were identified as Achromobacter insolitus and Zoogloea ramigera, respectively. A greenhouse experiment involving a non-sterilized soil was carried out with the aim to study the endophytic feature of these strains. After 40 days from inoculation, all the strains were in the inner of roots, but they were not detected in soil. In order to assess the location inside wheat plants, an experiment was conducted under axenic conditions. Fifteen days after inoculation, preparations of inoculated plants were observed by the scanning electron microscope, using the cryofracture technique, and by the transmission electron microscope. It was observed that all isolates were present on the external part of the roots and in the inner part at the elongation region, in cortex cells, but not in the endodermis or in the vascular bundle region. No colonizing bacterial cells were observed in wheat leaves.     

Salt tolerance in <Emphasis Type="Italic">Zea mays</Emphasis> (L). following inoculation with <Emphasis Type="Italic">Rhizobium</Emphasis> and <Emphasis Type="Italic">Pseudomonas</Emphasis>

This study aimed to investigate the effect of inoculation with plant growth-promoting Rhizobium and Pseudomonas species on NaCl-affected maize. Two cultivars of maize (cv. Agaiti 2002 and cv. Av 4001) selected on the basis of their yield potential were grown in pots outdoors under natural conditions during July. Microorganisms were applied at seedling stage and salt stress was induced 21 days after sowing and maintained up to 50% flowering after 120 days of stress. The salt treatment caused a detrimental effect on growth and development of plants. Co-inoculation resulted in some positive adaptative responses of maize plants under salinity. The salt tolerance from inoculation was generally mediated by decreases in electrolyte leakage and in osmotic potential, an increase in osmoregulant (proline) production, maintenance of relative water content of leaves, and selective uptake of K ions. Generally, the microbial strain acted synergistically. However, under unstressed conditions, Rhizobium was more effective than Pseudomonas but under salt stress the favorable effect was observed even if some exceptions were also observed. The maize cv. Agaiti 2002 appeared to be more responsive to inoculation and was relatively less tolerant to salt compared to that of cv. Av 4001.     

Genetic and bioclimatic variation in <Emphasis Type="Italic">Solanum pimpinellifolium</Emphasis>

Solanum pimpinellifolium, due to its close relationship to S. lycopersicum, has been a genetic source for many commercially important tomato traits. It is a wild species found in the coastal areas of Peru and Ecuador. In this study, the genetic variation of S. pimpinellifolium was studied using the diversity found in 10 microsatellites in 248 plants spread throughout its entire distribution area, including Ecuador, which has been underrepresented in previous studies. Peruvian and Ecuadorian accessions are genetically quite differentiated. A possible cause of these differences could be the non-uniform nature of the coastal Ecuadorian and Peruvian climates, seeing as an important correlation between genetic differentiation and climate has been found. In addition, Ecuadorian and south Peruvian accessions have a lower genetic diversity and a higher homozygosity due to their higher autogamy, lower population size, and possible colonization bottlenecks. The Galápagos Islands population is an extreme case, with no diversity, likely caused by a recent colonization from the northern continental Ecuadorian region where genetically identical plants have been found. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Elena Zuriaga and José M. Blanca contributed equally to this work.     

The Distribution of <Emphasis Type="Italic">Perilla</Emphasis> Species

Perilla (Lamiaceae) contains one tetraploid species, P. frutescens (L.) Britt. and three diploid species, P. citriodora (Makino) Nakai, P. hirtella Nakai and P. setoyensis G. Honda. Tetraploid species have been traditionally cultivated in Asia for their seed oil and for their fragrant leaves that are used as medicine or as a garnish for fish. The center of diversity is still obscure. To conserve the genetic resources, it is important to know the diversity of the tetraploid species. The three diploid species, which are possible parents of the tetraploid species, are all believed to be indigenous to Japan. Their distribution in China and Korea was clarified on the basis of herbarium and field surveys. The tetraploid species is assumed to have originated somewhere around the mid-to downstream area of the Changjiang River. Though Perilla is not cultivated as often in these areas as in northern China, Korea, the Himalayan region, or Myanmar, these areas should also be important for the conservation of genetic resources of tetraploid Perilla crops because of the expected high genetic diversity.     

<Emphasis Type="Italic">Bacillus subtilis</Emphasis> SQR 9 can control <Emphasis Type="Italic">Fusarium</Emphasis> wilt in cucumber by colonizing plant roots

Fusarium wilt is one of the major constraints on cucumber production worldwide. Several strategies have been used to control the causative pathogen, Fusarium oxysporum f. sp. cucumerinum J. H. Owen, including soil solarization, fungicide seed treatment and biological control. In this study, F. oxysporum f. sp. cucumerinum was successfully controlled by a newly isolated strain, Bacillus subtilis SQR 9, in vitro and in vivo. Greenhouse experiments were carried out to evaluate the effect of inoculation and solid fermentation of organic fertilizer with B. subtilis SQR 9, hereby defined as bio-organic fertilizer (BIO), on the control of Fusarium wilt. In comparison with the control, the wilt incidence was significantly reduced (49–61% reduction) by application of BIO. The rhizosphere population of F. oxysporum f. sp. cucumerinum, as detected both by selective plating and realtime PCR, was significantly lower in BIO-treated plants than the control. The localization of bacterial cells, pattern of colonization and survival of B. subtilis SQR 9 in the rhizsosphere of cucumber, was examined by fluorescent microscopy and explored following recovery of the green fluorescent protein (gfp)-labeled SQR 9 with the new gfp-marked shuttle vector pHAPII through selective plating. The preferential sites of the labeled strain were the differentiation and elongation zone, root hair and the lateral root junctions. The population of the strain was 10⁶ cfu/g root in rhizoplane. These results indicate that the strain was able to survive well in the rhizosphere of cucumber, suppressed growth of F. oxysporum in the rhizosphere of cucumber and protected the host from the pathogen.     

Evaluation and utilization of <Emphasis Type="Italic">Aegilops</Emphasis> and wild <Emphasis Type="Italic">Triticum</Emphasis> species for enhancing iron and zinc content in wheat

Grains of 80 accessions of nine species of wild Triticum and Aegilops along with 15 semi-dwarf cultivars of bread and durum wheat grown over 2 years at Indian Institute of Technology, Roorkee, were analyzed for grain iron and zinc content. The bread and durum cultivars had very low content and little variability for both of these micronutrients. The related non-progenitor wild species with S, U and M genomes showed up to 3–4 folds higher iron and zinc content in their grains as compared to bread and durum wheat. For confirmation, two Ae. kotschyi Boiss. accessions were analyzed after ashing and were found to have more than 30% higher grain ash content than the wheat cultivars containing more than 75% higher iron and 60% higher zinc than that of wheat. There were highly significant differences for iron and zinc contents among various cultivars and wild relatives over both the years with very high broad sense heritability. There was a significantly high positive correlation between flag leaf iron and grain iron (r = 0.82) and flag leaf zinc and grain zinc (r = 0.92) content of the selected donors suggesting that the leaf analysis could be used for early selection for high iron and zinc content. ‘Chinese Spring’ (Ph ^I ) was used for inducing homoeologous chromosome pairing between Aegilops and wheat genomes and transferring these useful traits from the wild species to the elite wheat cultivars. A majority of the interspecific hybrids had higher leaf iron and zinc content than their wheat parents and equivalent or higher content than their Aegilops parents suggesting that the parental Aegilops donors possess a more efficient system for uptake and translocation of the micronutrients which could ultimately be utilized for wheat grain biofortification. Partially fertile to sterile BC₁ derivatives with variable chromosomes of Aegilops species had also higher leaf iron and zinc content confirming the possibility of transfer of required variability. Some of the fertile BC₁F₃ and BC₂F₂ derivatives had as high grain ash and grain ash iron and zinc content as that of the donor Aegilops parent. Further work on backcrossing, selfing, selection of fertile derivatives, leaf and grain analyses for iron and zinc for developing biofortified bread and durum wheat cultivars is in progress. Nidhi Rawat, Vijay K. Tiwari, and Neelam Singh have contributed equally to the work.     

<Emphasis Type="Italic">Rhizobium</Emphasis>,<Emphasis Type="Italic"> Bradyrhizobium</Emphasis> and<Emphasis Type="Italic"> Agrobacterium</Emphasis> strains isolated from cultivated legumes

The present study was conducted to isolate and characterize rhizobial strains from root nodules of cultivated legumes, i.e. chickpea, mungbean, pea and siratro. Preliminary characterization of these isolates was done on the basis of plant infectivity test, acetylene reduction assay, C-source utilization, phosphate solubilization, phytohormones and polysaccharide production. The plant infectivity test and acetylene reduction assay showed effective root nodule formation by all the isolates on their respective hosts, except for chickpea isolate Ca-18 that failed to infect its original host. All strains showed homology to a typical Rhizobium strain on the basis of growth pattern, C-source utilization and polysaccharide production. The strain Ca-18 was characterized by its phosphate solubilization and indole acetic acid (IAA) production. The genetic relationship of the six rhizobial strains was carried out by random amplified polymorphic DNA (RAPD) including a reference strain of Bradyrhizobium japonicum TAL-102. Analysis conducted with 60 primers discriminated between the strains of Rhizobium and Bradyrhizobium in two different clusters. One of the primers, OPB-5, yielded a unique RAPD pattern for the six strains and well discriminated the non-nodulating chickpea isolate Ca-18 from all the other nodulating rhizobial strains. Isolate Ca-18 showed the least homology of 15% and 18% with Rhizobium and Bradyrhizobium, respectively, and was probably not a (Brady)rhizobium strain. Partial 16S rRNA gene sequence analysis for MN-S, TAL-102 and Ca-18 strains showed 97% homology between MN-S and TAL-102 strains, supporting the view that they were strains of B. japonicum species. The non-infective isolate Ca-18 was 67% different from the other two strains and probably was an Agrobacterium strain.     

Arbuscular mycorrhizal fungi mediated uptake of nutrient elements by Chinese milk vetch (<Emphasis Type="Italic">Astragalus sinicus</Emphasis> L.) grown in lanthanum spiked soil

A greenhouse experiment was conducted to study the effect of mycorrhizal colonization by Gigaspora margarita, Glomus intraradices, and Acaulospora laevis on nutrient uptake of K, Ca, Mg, Cu, Zn, Fe, and Mn by Astragalus sinicus L. in soils spiked with lanthanum at five rates (0, 1, 5, 10, and 20 mg kg⁻¹). Lanthanum application significantly decreased the concentrations of K, Ca, Mg, Cu, Zn, and Fe in shoots and the concentrations of Cu and Zn in roots. Mycorrhizal treatments markedly improved uptake of nutrients, and these results are important since nutrient deficiency often occurs in contaminated sites.