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.     

Effects of an arbuscular mycorrhizal fungus and two plant-parasitic nematodes on<Emphasis Type="Italic"> Musa</Emphasis> genotypes differing in root morphology

In this study, the effect of an arbuscular mycorrhizal fungus (AMF) and two migratory endoparasitic nematodes on Musa plant growth, including the root system, were examined. In addition, the AMF-nematode interaction was studied. Seven Musa genotypes with different root systems were selected. Based on their relative mycorrhizal dependency, two genotypes (Calcutta 4 and Obino l'Ewai) were selected for AMF-nematode interaction studies. The experiments were performed under greenhouse conditions. Mycorrhization with Glomus mosseae resulted in a significantly better plant growth even in the presence of nematodes. The effect of AMF on the root system was genotype-dependent and seemed to be related to the relative mycorrhizal dependency of the genotype. The nematodes also affected the root system, decreasing branching. Nematode population densities were significantly reduced in the presence of AMF, except for Pratylenchus coffeae in Obino l'Ewai. In the root system, it appeared that the decreased branching caused by the nematodes was counterbalanced by the increased branching caused by the AMF.     

The effect of rhizobiophages on<Emphasis Type="Italic"> Sinorhizobium meliloti</Emphasis> -<Emphasis Type="Italic"> Medicago sativa</Emphasis> symbiosis

Using Tn5 mutagenesis of lysogenic Sinorhizobium meliloti strain L5–30 (L54), three neomycin-resistant transconjugants differing in phage-resistance profiles were isolated. Two of them increased the dry weight of plants and were capable of establishing root nodules, whereas the third one was ineffective. The bacterium-phage interaction did not have observable consequences in the Medicago sativa - S. meliloti symbiosis because it did not affect the number of nodules on M. sativa or plant dry weight.     

A new mycorrhizal helper bacterium, <Emphasis Type="Italic">Ralstonia</Emphasis> species,in the ectomycorrhizal symbiosis between <Emphasis Type="Italic">Pinus thunbergii</Emphasis> and <Emphasis Type="Italic">Suillus granulatus</Emphasis>

In a Robinia-pseudoacacia-dominated coastal forest in Tottori prefecture Japan, the growth and survival of Pinus thunbergii seedlings and the natural regeneration of P. thunbergii was disturbed by R. pseudoacacia. In order to improve the growth of P. thunbergii seedling in the Tottori sand dune, we tried to find a mycorrhiza helper bacteria (MHB) from P. thunbergii mycorrhizosphere in a Tottori sand dune. Two MHB, Ralstonia sp. and Bacillus subtilis, were selected from the nine bacterial species isolated from the mycorrhizosphere of P. thunbergii. The bacterial effect on the ectomycorrhizal fungus Suillus granulatus was investigated by confrontation assay and a microcosm experiment. The confrontation assay showed that Ralstonia sp. promoted the hyphal growth of S. granulatus. Moreover, the S. granulatus–P. thunbergii symbiosis was significantly stimulated by Ralstonia sp. and B. subtilis. Ralstonia sp. and B. subtilis were regarded as MHB associated with P. thunbergii. This is the first report of Ralstonia sp. as an MHB.     

Characterization of rhizobia from <Emphasis Type="Italic">Sesbania</Emphasis> species native to seasonally wetland areas in Uruguay

Naturally growing Sesbania species with tolerance to unfavourable habitats are widely distributed in non-cultivated seasonally wetland areas in Uruguay. We investigated the relative abundance, diversity and symbiotic efficiency of Sesbania punicea and S. virgata rhizobia in three ecologically different undisturbed and water-logged sites in Uruguay. Numbers of native-soil rhizobia infective on S. punicea or S. virgata were low, with higher numbers associated with the presence of S. virgata. Plants of S. virgata inoculated with soil suspension showed aerial and nodule biomass greater than that obtained with S. punicea. The rhizobia nodulating Sesbania species in water-logged lands in different regions of Uruguay were diverse differing in growth rates, acid production, growth at 39°C and in LB medium, host range and symbiotic efficiency. Seventeen representative strains clustered into four groups on the basis of phenotypic characteristics, ARDRA and DNA fingerprinting (GTG₅-PCR). Partial sequence of 16S rRNA from eight of these strains classified them into at least two genera with four species: Azorhizobium doebereinerae, Rhizobium sp. related to R. etli and two different Rhizobium sp.-Agrobacterium. Our results confirm the presence of the specie Azorhizobium doebereinerae as microsymbionts of S. virgata in South America. No strain of Rhizobium etli has previously been reported as a microsymbiont of Sesbania, though R. etli like organisms have also been recovered from Dalea purpurea and Desmanthus illinoensis. Significant increases in dry matter production were obtained with S. virgata plants inoculated with selected rhizobial strains under growth chamber conditions.     

<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.     

Differences in native soil ecology associated with invasion of the exotic annual chenopod,<Emphasis Type="Italic"> Halogeton glomeratus</Emphasis>

Various biotic and abiotic components of soil ecology differed significantly across an area where Halogeton glomeratus is invading a native winterfat, [ Krascheninnikovia (= Ceratoides) lanata] community. Nutrient levels were significantly different among the native, ecotone, and exotic-derived soils. NO₃, P, K, and Na all increased as the cover of halogeton increased. Only Ca was highest in the winterfat area. A principal components analysis, conducted separately for water-soluble and exchangeable cations, revealed clear separation between halogeton- and winterfat-derived soils. The diversity of soil bacteria was highest in the exotic, intermediate in the ecotone, and lowest in the native community. Although further studies are necessary, our results offer evidence that invasion by halogeton alters soil chemistry and soil ecology, possibly creating conditions that favor halogeton over native plants.     

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.     

Genetic Characterization of Brazilian Annual <Emphasis Type="Italic">Arachis</Emphasis> Species from Sections <Emphasis Type="Italic">Arachis</Emphasis> and <Emphasis Type="Italic">Heteranthae</Emphasis> using RAPD Markers

The genus Arachis is divided into nine taxonomic sections. Section Arachis is composed of annual and perennial species, while section Heteranthae has only annual species. The objective of this study was to investigate the genetic relationships among 15 Brazilian annual accessions from Arachis and Heteranthae using RAPD markers. Twenty-seven primers were tested, of which nine produced unique fingerprintings for all the accessions studied. A total of 88 polymorphic fragments were scored and the number of fragments per primer varied from 6 to 17 with a mean of 9.8. Two specific markers were identified for species with 2n = 18 chromosomes. The phenogram derived from the RAPD data corroborated the morphological classification. The bootstrap analysis divided the genotypes into two significant clusters. The first cluster contained all the section Arachis species, and the accessions within it were grouped based upon the presence or absence of the ‘A’ pair and the number of chromosomes. The second cluster grouped all accessions belonging to section Heteranthae.     

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.     

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.     

Phosphate solubilization by <Emphasis Type="Italic">Penicillium</Emphasis> spp. closely associated with wheat roots

In this study we found that Penicillium spp. exhibiting P-solubilizing activity are common both on and in the roots of wheat plants grown in southern Australian agricultural soils. From 2,500 segments of washed and surface-disinfested root pieces, 608 and 223 fungi were isolated on a selective medium, respectively. All isolates were screened for P solubilization on solid medium containing hydroxyapatite (HA); 47 isolates (5.7%) solubilized HA and were identified as isolates of Penicillium or its teleomorphs. These isolates were evaluated for solubilization of Idaho rock phosphate (RP) in liquid culture. Penicillium bilaiae strain RS7B-SD1 was the most effective, mobilizing 101.7 mg P l^–1 after 7 days. Other effective isolates included Penicillium simplicissimum (58.8 mg P l^–1), five strains of Penicillium griseofulvum (56.1–47.6 mg P l^–1), Talaromyces flavus (48.6 mg P l^–1) and two unidentified Penicillium spp. (50.7 and 50 mg P l^–1). A newly isolated strain of Penicillium radicum (KC1-SD1) mobilized 43.3 mg P l^–1. RP solubilization, biomass production and solution pH for P. bilaiae RS7B-SD1, P. radicum FRR4718 or Penicillium sp. 1 KC6-W2 was determined over time. P. bilaiae RS7B-SD1 solubilized the greatest amount of RP (112.7 mg P l^–1) and had the highest RP-solubilizing activity per unit of biomass produced (up to 603.2 g P l^–1 mg biomass^–1 at 7 days growth). This study has identified new isolates of Penicillium fungi with high mineral phosphate solubilizing activity. These fungi are being investigated for the ability to increase crop production on strong P-retaining soils in Australia.     

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.     

Genetic diversity of the D-genome in <Emphasis Type="Italic">T. aestivum</Emphasis> and <Emphasis Type="Italic">Aegilops</Emphasis> species using SSR markers

Simple sequence repeats (SSRs), highly dispersed nucleotide sequences in genomes, were used for germplasm analysis and estimation of the genetic relationship of the D-genome among 52 accessions of T. aestivum (AABBDD), Ae. tauschii (D^tD^t), Ae. cylindrica (CCD^cD^c) and Ae. crassa (MMD^cr1D^cr1), collected from 13 different sites in Iran. A set of 21 microsatellite primers, from various locations on the seven D-genome chromosomes, revealed a high level of polymorphism. A total of 273 alleles were detected across all four species and the number of alleles per each microsatellite marker varied from 3 to 27. The highest genetic diversity occurred in Ae. tauschii followed by Ae. crassa, and the genetic distance was the smallest between Ae. tauschii and Ae. cylindrica. Data obtained in this study supports the view that genetic variability in the D-genome of hexaploid wheat is less than in Ae. tauschii. The highest number of unique alleles was observed within Ae. crassa accessions, indicating this species as a great potential source of novel genes for bread wheat improvement. Knowledge of genetic diversity in Aegilops species provides different levels of information which is important in the management of germplasm resources.     

<Emphasis Type="Italic">Calamintha nepeta</Emphasis> (L.) Savi and <Emphasis Type="Italic">Micromeria thymifolia</Emphasis> (Scop.) Fritsch cultivated in Italy

Calamintha nepeta and Micromeria thymifolia have been traditionally used in the Mediterranean area as condiments and medicinal plants for a long time. Whereas in parts of Italy C. nepeta (special recipes have been developed in Lazio and Tuscany) is also an established garden plant showing different evolutionary products and their interaction among each other and the wild progenitor, M. thymifolia is being developed into a new crop plant. Both plants and their uses are described with regard to Italy. There is a marked tendency to broaden the use of condiments and spices which results in new crop plants which have to be documented and elaborated in further studies. Many species of Labiatae are predisposed to use by man and new items can be found even in areas which have to be considered as well studied.     

Field inoculation effectiveness of native and exotic arbuscular mycorrhizal fungi in a Mediterranean agricultural soil

In sustainable agriculture, arbuscular mycorrhizal (AM) fungal inoculation in agronomical management might be very important, especially when the efficiency of native inocula is poor. Here, we assessed the effect of native and exotic selected AM fungal inocula on plant growth and nutrient uptake in a low input Trifolium alexandrinum-Zea mays crop rotation. We evaluated the effects of four exotic AM fungal isolates on T. alexandrinum physiological traits in greenhouse. Then, the field performances of T. alexandrinum inoculated with the exotic AMF, both single and mixed, were compared to those obtained with a native inoculum, using a multivariate analysis approach. Finally, we tested the residual effect of AM fungal field inoculation on maize as following crop. Multivariate analysis showed that the field AM fungal inoculation increased T. alexandrinum and Z. mays productivity and quality and that the native inoculum was as effective as, or more effective than, exotic AM fungal isolates. Moreover, the beneficial effects of AMF were persistent until the second year after inoculation. The use of native AMF, produced on farm with mycotrophic plants species, may represent a convenient alternative to commercial AM fungal inocula, and may offer economically and ecologically important advantages in sustainable or organic cropping systems.     

AFLP and RFLP linkage map in <Emphasis Type="Italic">Coix</Emphasis>

Coix is a genus in the grass family placed in the tribe Maydeae. It is closely related to maize and is also used as a crop plant. Since many valuable traits have been identified recently in Coix, it is considered to be a valuable genetic resource, particularly for maize improvement. In this study, a Coix genetic linkage map was constructed using an F2 population of 131 individuals. Eighty AFLP and 10 RFLP markers were mapped, covering a total length of 1339.5 cM with an average interval of 14.88 cM. The map consisted of 10 linkage groups, were consistent with the chromosome numbers observed cytogenetically. Both AFLP and RFLP markers were used first for genetic analysis in Coix. AFLP markers were generated by two restriction enzyme combinations, EcoRI/MseI and PstI/MseI. A total of 1349 bands were amplified, of which 140 were polymorphic. The polymorphism detection efficiency of the two enzyme combinations was compared, and utility of AFLP markers to construct the linkage map was discussed. Ten RFLP markers detected by three different probes were distributed on eight different linkage groups. The results provide a foundation to map and isolate important genes in Coix, and to investigate its genomic architecture, possible origins, and relationship with maize at the DNA level.     

A bradyrhizobial-<Emphasis Type="Italic">Penicillium</Emphasis> spp. biofilm with nitrogenase activity improves N<Subscript>2</Subscript> fixing symbiosis of soybean

A bradyrhizobial-fungal biofilm (i.e. Bradyrhizobium elkanii SEMIA 5019-Penicillium spp.) developed in vitro was assayed for its nitrogenase activity and was evaluated for N₂-fixing symbiosis with soybean under greenhouse conditions. The biofilm showed nitrogenase activity, but the bradyrhizobial strain alone did not. Shoot and root growth, nodulation and N accumulation of soybean increased significantly with an inoculum developed from the biofilm. This study concludes that such biofilmed inoculants can improve N₂-fixing symbiosis in legumes, and can also directly contribute to soil N fertility in the long term. Further studies should be conducted to investigate the performance of these inoculants under field conditions.     

Identification of <Emphasis Type="Italic">Aegilops</Emphasis> L. species and <Emphasis Type="Italic">Triticum aestivum</Emphasis> L. based on chloroplast DNA

The genus Aegilops L. is a very important genetic resource for the breeding of bread wheat Triticum aestivum. Therefore, an accurate and easy identification of Aegilops species is required. Traditionally, identification of Aegilops species has relied heavily on morphological characters. These characters, however, are either not variable enough among Aegilops species or too plastic to be used for identification at the species level. Molecular markers that are more stable within species, therefore, could be the alternative strategy towards an accurate identification. Since the chloroplast DNA has a lower level of evolution compared to the nuclear genome, an attempt was made in this study to investigate polymorphism in the chloroplast DNA among 21 Aegilops species (including Ae. mutica that is now known as Amblyopyrum muticum) and between the latter and T. aestivum to generate markers for the diagnosis of all targeted species. Cleaved amplified polymorphic sequence (CAPS) applied on 22 coding and non-coding chloroplast regions using 80 endonucleases and sequencing of two of those regions revealed little polymorphism between T. aestivum and the various Aegilops species examined and to a less extent was the variation among Aegilops species. Polymorphism observed among species analysed allowed the discrimination of T. aestivum and 12 Aegilops species.