Growth and Nutrition of Eucalypt Rooted Cuttings Promoted by Ectomycorrhizal Fungi in Commercial Nurseries

ABSTRACT Ectomycorrhizal fungi (EMF) may improve the adaptation of eucalypts saplings to field conditions and allow more efficient fertilizer use. The effectiveness of EMF inoculum application in promoting fungal colonization, plant growth, nutrient uptake, and the quality of rooted cuttings was evaluated forEucalyptus urophylla under commercial nursery conditions. For inoculated treatments, fertilization of the sapling substrate was reduced by 50 %. The experiment was carried out in a completely randomized design in a 4 × 4 factorial arrangement, wherein the factors were inoculum application rates of 0 (control), 5, 10, and 15 gel beads of calcium alginate containing the vegetative mycelium of Amanita muscaria, Elaphomyces antracinus, Pisolithus microcarpus, andScleroderma areolatum, plus a non-inoculated treatment without fertilization reduction in the substrate (commercial). Ectomycorrhizal fungi increased plant growth and fungal colonization as well as N and K uptake evenly. The best plant growth and fungal colonization were observed for the highest application rate. The greatest growth and fungal colonization and contents of P, N, and K were observed at the 10-bead rate. Plant inoculation with Amanita muscaria, Elaphomyces anthracinus, and Scleroderma areolatum increased P concentrations and contents in a differential manner. The Dickson Quality Index was not affected by the type of fungi or by inoculum application rates. Eucalypt rooted cuttings inoculated with ectomycorrhizal fungi and under half the amount of commercial fertilization had P, N, and K concentrations and contents greater than or equal to those of commercial plants and have high enough quality to be transplanted after 90 days.

Rhizobia Isolated from Coal Mining Areas in the Nodulation and Growth of Leguminous Trees

ABSTRACT An alternative for recovery of areas degraded by coal mining is revegetation with rapidly growing leguminous trees, which often do not establish in low fertility soils. The objective of this study was to evaluate the efficiency of native rhizobia isolated from coal mining areas in the nodulation and growth of leguminous trees. We isolated 19 strains of rhizobia from a degraded soil near Criciúma, SC, Brazil, and evaluated the nodulation and growth-promoting capacity of the inoculated isolates for bracatinga (Mimosa scabrella), maricá (M. bimucronata) and angico-vermelho (Parapiptadenia rigida). Isolates UFSC-B2, B6, B8, B9, B11 and B16 were able to nodulate bracatinga, providing average increases of 165 % in shoot dry matter, with a significant contribution to N accumulation. Isolates UFSC-B5, B12, and M8 favored nodulation and growth of maricá, especially isolate UFSC-B12, which promoted increases of 370 % in N accumulation compared to treatment with N fertilizer. All strains were inefficient in promoting growth and N uptake by angico-vermelho. In conclusion, isolation and use of selected rhizobia for bracatinga and maricá plant inoculation can contribute to the growth and accumulation of N, with prospects for use in programs for revegetation of degraded soils in coal mining areas.

Nitrogen translocation in wheat inoculated with Azospirillum and fertilized with nitrogen

The productivity and the translocation of assimilates and nitrogen (N) were compared after inoculation of wheat (Triticum aestivum L., cv. BR-23) seeds with two strains of Azospirillum brasilense (strains 245 and JA 04) under field conditions. The inoculation of wheat seeds was done with a peat inoculant at sowing time. Plant material for evaluations were collected at anthesis and maturity. No differences in grain yield and in the translocation of assimilates resulting from inoculation were detected. Differences were observed in relation to N rates (0, 15, and 60 kg ha-1). N content in the grain increased significantly in the bacteria-inoculated treatments in which N was not added. This increase in N content in the grain with inoculation was probably due to higher N uptake after anthesis without any significant contribution on the grain yield. Such increment was of 8.4 kg ha-1 of N representing 66% more N than in no inoculated treatment. Regardless of the inoculation and the rate of N applied, it was observed that about 70% of the N accumulated at anthesis was translocated from vegetative parts to the grain.

Preservation of rhizobia by lyophilization with trehalose

The objective of this study was to determine if trehalose can effectively enhance the viability and storage lifetime of Rhizobium tropici and Rhizobium etli after lyophilization as compared to the traditional protective combination of sucrose and peptone. Two strains of bean-nodulating rhizobia were effectively preserved in the lyophilized state by 100 mM trehalose, and survived for at least 12 days, even when stored under non-ideal conditions. Trehalose provided better protection than the sucrose/peptone combination. When protective excipients were introduced inside the cells, strain CFN 42 was more sensitive to the type of additive used than strain CIAT 899, probably because CIAT 899 produces beta (1-2) glucan, which may have a protective effect. beta (1-2) glucan combined with sucrose protected better than trehalose against leakage from freeze-dried liposomes, but by itself it had no protective capability. There was a high level of unfrozen water associated with the hydrated glucan, in addition to a thermal event at approximately 70ºC, corresponding to an observed gel-sol transition in the glucan. These findings may shed light on the mechanism by which the glucan contributes to the protection of dried cells and liposomes, and may lead to further improvements in rhizobial inoculants.

Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orange

The development and optimization of efficient transformation protocols is essential in new citrus breeding programs, not only for rootstock, but also for scion improvement. Transgenic 'Hamlin' sweet orange (Citrus sinensis (L.) Osbeck) plants were obtained by Agrobacterium tumefaciens-mediated transformation of epicotyl segments collected from seedlings germinated in vitro. Factors influencing genetic transformation efficiency were evaluated including seedling incubation conditions, time of inoculation with Agrobacterium and co-culture conditions. Epicotyl segments were adequate explants for transformation, regenerating plants by direct organogenesis. Higher percentage of transformation was obtained with explants collected from seedlings germinated in darkness, transferred to 16 hours photoperiod for 2-3 weeks, and inoculated with Agrobacterium for 15-45 min. The best co-culture condition was the incubation of the explants in darkness, for three days in culture medium supplemented with 100 muM of acetosyringone. Genetic transformation was confirmed by performing beta-glucoronidase (GUS) assays and, subsequently, by PCR amplification for the nptII and GUS genes.

Resistance spectra of six elite breeding lines of upland rice to Pyricularia grisea

The objective of this work was to evaluate the resistance spectra of six elite breeding lines of rice, developed for improved yield and grain quality, in inoculation tests in the greenhouse and in the field. Forty-six isolates of Pyricularia grisea collected from the cultivar Primavera, 31 from the cultivar Maravilha and 19 from six elite breeding lines, totaling 96 were utilized for inoculations. Out of 11 international and 15 Brazilian pathotypes, IC-1, IB-9, and BD-16, respectively, were identified as most frequent isolates collected from the cultivar Primavera. The isolates retrieved from Maravilha belong to four international and 11 Brazilian pathotypes, the predominant ones being IB-9 and IB-49 and BB-1 and BB-21, respectively. Lines CNAs 8711 and CNAs 8983 showed resistant reaction to all test isolates from Maravilha, while CNAs 8983 was susceptible to three isolates of Primavera pertaining to the pathotype IC-1. A majority of isolates exhibiting compatible reaction to Primavera were incompatible to Maravilha and vice-versa.Field assessment of rice blast utilizing the area under disease progress curve as a criterion for measuring disease severity showed significant differences among the six breeding lines. The isolates of P. grisea exhibiting differential reaction on breeding lines can be utilized in pyramiding resistance genes in new upland rice cultivars.

Diallel analysis for frogeye leaf spot resistance in soybean

Seven soybean cultivars (Bossier, Cristalina, Davis, Kent, Lincoln, Paraná and Uberaba), with different levels of resistance to Cercospora sojina, were crossed in a diallel design to determine the general (GCA) and specific (SCA) combining abilities relative to the inheritance of the resistance. Race 04 of the fungus was inoculated in the parents and in the 21 F1 hybrids in a greenhouse in a completely randomized design, with 12 replications. The reactions to the disease were evaluated 20 days after the inoculation, always on the most infected leaflet. Both GCA and SCA were significant for all the evaluated characters, being inferred that, for the expression of the characters, the additive, dominant and, possibly, epistatic genic actions were important. The largest values of estimated SCA effect (ij) were observed in the hybrid combinations where at least one parent presented high GCA. Cristalina, Davis and Uberaba cultivars showed the largest estimates for GCA effect (i), and from the analysis of ii, the contribution of these parents to heterosis of their hybrids will be towards the reduction of the disease symptoms. Therefore, these cultivars are indicated as parents in breeding programs that seek the development of soybean cultivars with resistance to frogeye leaf spot.

Lipoxygenase activities during development of root and nodule of soybean

The objective of this work was to evaluate root and nodule soybean lipoxygenases in Doko cultivar and in a near isogenic line lacking seed lipoxygenases, inoculated and uninoculated with Bradyrhizobium elkanii. The lipoxygenase activities from roots collected at 3, 5, 9, 13, 18 and 28 days post-inoculation and from nodules collected at 13, 18 and 28 days post-inoculation were measured. The pH-activity profiles from root and nodules suggested that the lipoxygenases pool expressed in these organs from Doko cultivar and triple-null near isogenic lines are similar. The root lipoxygenase activity of Doko and triple-null lines, inoculated and uninoculated, reduced over time. The highest lipoxygenase activity observed at the beginning of root formation suggests the involvement of this enzyme in growth and development of this organ. However, for nodules an expressive increase of lipoxygenase activity was noticed 28 days post-inoculation. Root and nodule showed, at least, two mobility groups for lipoxygenases in immunoblottings, with approximately 94 and 97 kDa.

Reaction of arracacha genotypes to the root soft rot caused by Pectobacterium chrysanthemi

The purpose of this paper was to screen thirty-two arracacha genotypes for their reaction to root soft rot. Twenty roots of each genotype were inoculated with two Pectobacterium chrysanthemi isolates in a randomized experiment (10 roots/isolate). After inoculation, roots were individually wrapped with PVC film and kept at 26ºC in closed plastic bags. Soft rot lesions were recorded after 36 hours and genotypes were grouped in four classes of susceptibility by cluster analysis: 10 were less susceptible, 16 intermediate, 3 susceptible and 3 very susceptible. All the tested arracacha genotypes showed only variation in the degree of susceptibility.

Performance of Holstein heifers fed sugarcane silages treated with urea, sodium benzoate or Lactobacillus buchneri

The objective of this work was to evaluate the performance of heifers fed sugarcane silages produced with and without additives. Thirty-two Holstein heifers were randomly assigned, in a block design, to evaluate rations (46% silage; 54% concentrate; 12% crude protein) containing silages treated with (fresh basis) urea (0.5%), sodium benzoate (0.1%) or Lactobacillus buchneri (3.64x10(5) cfu g-1 ). Inoculation with L. buchneri improved daily gain (1.24 vs. 0.94 kg day-1 ), and the addition of benzoate resulted in better feed conversion (7.6 vs. 9.4 kg of dry matter per kg of live weight), in relation to the untreated silage (control). Treatments did not affect dry matter intake (mean of 2.19% of live weight). Rations containing silages treated with benzoate or L. buchneri showed lower cost per kg of weight gain. Treatment with urea did not improve animal performance, but the cost per kg of weight gain was lower than that of the control ration.

Characterization of Tomato yellow spot virus, a novel tomato-infecting begomovirus in Brazil

The objective of this work was the biological and molecular characterization of a begomovirus detected in São Joaquim de Bicas, Minas Gerais, Brazil, named TGV-[Bi2], by determining its host range, complete nucleotide sequence and phylogenetic relationships with other begomoviruses. Biological characterization consisted of a host range study using either sap inoculation or particle bombardment as inoculation methods. The yellow spot virus can infect plants in Solanaceae and Amaranthaceae, including economically importat crops as sweet pepper, and weeds as Datura stramonium and Nicotiana silvestris. For the molecular characterization, the full-length genome (DNA-A and DNA-B) was amplified, cloned and completely sequenced. Sequence comparisons and phylogenetic analyses indicated that TGV-[Bi2] constitutes a novel begomovirus species named Tomato yellow spot virus (ToYSV), closely related to Sida mottle virus (SiMoV).

New isolates of ectomycorrhizal fungi and the growth of eucalypt

The objective of this work was to evaluate the efficiency of ectomycorrhizal isolates on root colonization, phosphorus uptake and growth of Eucalyptus dunnii seedlings. Inocula of ten ectomycorrhizal isolates of Chondrogaster angustisporus, Hysterangium gardneri, Pisolithus spp., and Scleroderma spp. were aseptically produced in a peat-vermiculite mixture supplemented with liquid culture medium. Plants grew in a similar substrate supplemented with macro-and micro-nutrients; treatments were randomly distributed in a greenhouse. After three months, seedlings inoculated with three isolates - UFSC-Sc68 (Scleroderma sp.), UFSC-Ch163 (Chondrogaster angustisporus), and UFSC-Pt188 (Pisolithus microcarpus) - had a phosphorus shoot content and a shoot dry matter higher or equivalent to those of noninoculated controls which had been fertilized with a 16-fold phosphorus amount. These isolates were selected for new studies for establishing inoculum production techniques, in order to be applied in reforestation programmes under nursery and field conditions.

Inheritance of resistance to cotton blue disease

The objective of this work was to determine the inheritance of cotton blue disease resistance by cotton plants. Populations derived from the CD 401 and Delta Opal resistant varieties were evaluated, through a greenhouse test with artificial inoculation by viruliferous aphids. Cotton blue disease resistance is conditioned by one dominant gene, both in CD 401 and Delta Opal varieties.

Mycorrhizal dependency of mangaba tree under increasing phosphorus levels

The objective of this study was to evaluate the mycorrhizal dependency of mangaba tree (Hancornia speciosa) plantlets, under increasing levels of phosphorus fertilization. The experimental design was completely randomized in a 4×5 factorial arrangement with three mycorrhizal fungi inocula - Gigaspora margarita, Glomus etunicatum, or a pool of native mycorrhizal fungi (Acaulospora longula, Glomus clarum, Gigaspora albida, Paraglomus sp.) -, and a nonmycorrhizal control, in combination with five levels of phosphorus applied to the substrate: 0, 25, 50, 75, and 100 mg kg-1. After 180 days of growth, plantlets with inoculation of native mycorrhizal pool produced more shoot and root dry biomass and had higher shoot phosphorus content and accumulation. The noninoculated control showed the lowest values, independently of the phosphorus level. The highest relative mycorrhizal dependency occurred with the inoculation of native mycorrhizal fungi. Plants with mycorrhizal fungi did not respond to phosphorus addition above 50 mg kg-1. Mangaba tree is highly dependent on mycorrhiza, but the degree of dependency varies according to phosphorus levels and fungal inocula. In general, mangaba tree is more responsive to mycorrhizal fungi inoculation than to phosphorus addition.

Cell suspension culture and plant regeneration of a Brazilian plantain, cultivar Terra

The objective of this study was to establish cell suspension culture and plant regeneration via somatic embryogenesis of a Brazilian plantain, cultivar Terra Maranhão, AAB. Immature male flowers were used as explant source for generating highly embryogenic cultures 45 days after inoculation, which were used for establishment of cell suspension culture and multiplication of secondary somatic embryos. Five semisolid culture media were tested for differentiation, maturation, somatic embryos germination and for plant regeneration. An average of 558 plants per one milliliter of 5% SCV (settled cell volume) were regenerated in the MS medium, with 11.4 µM indolacetic acid and 2.2 µM 6-benzylaminopurine. Regenerated plants showed a normal development, and no visible somaclonal variation was observed in vitro. It is possible to regenerate plants from cell suspensions of plantain banana cultivar Terra using MS medium supplemented with 11.4 µM of IAA and 2.2 µM of BAP.