Rhizobacterial characterization for quality control of eucalyptus biogrowth promoter products

Abstract Plant growth-promoting rhizobacteria strains from special formulations have been used to optimize eucalyptus cutting production. To undertake quality control for the formulated products, the rhizobacterial strains should be characterized to assess their purity and authentication. In the present study, we characterized nine strains of rhizobacteria, including three Bacillus subtilis (S1, S2 and 3918), two Pseudomonas sp. (MF4 and FL2), P. putida (MF2), P. fulva (Ca), Frateuria aurantia (R1), and Stenotrophomonas maltophilia (CIIb). The strains were differentiated by colony morphology after 24 h of incubation in three different solid state culture media (glucose-nutritive agar, 523 medium and yeast extract-mannitol agar), sensitivity to a panel of 28 antibiotics (expressed according to the formation of inhibition halos of bacterial growth in the presence of antibiotics), and PCR-RFLP profiles of the 16S rDNA gene produced using nine restriction enzymes. It was possible to differentiate all nine strains of rhizobacteria using their morphological characteristics and sensitivity to antibiotics. The molecular analysis allowed us to separate the strains CIIb, FL2 and R1 from the strains belonging to the genera Bacillus and Pseudomonas. By using these three methods concomitantly, we were able to determine strain purity and perform the authentication.

Linkage analysis between dominant and co-dominant makers in full-sib families of out-breeding species

As high-throughput genomic tools, such as the DNA microarray platform, have lead to the development of novel genotyping procedures, such as Diversity Arrays Technology (DArT) and Single Nucleotide Polymorphisms (SNPs), it is likely that, in the future, high density linkage maps will be constructed from both dominant and co-dominant markers. Recently, a strictly genetic approach was described for estimating recombination frequency (r) between co-dominant markers in full-sib families. The complete set of maximum likelihood estimators for r in full-sib families was almost obtained, but unfortunately, one particular configuration involving dominant markers, segregating in a 3:1 ratio and co-dominant markers, was not considered. Here we add nine further estimators to the previously published set, thereby making it possible to cover all combinations of molecular markers with two to four alleles (without epistasis) in a full-sib family. This includes segregation in one or both parents, dominance and all linkage phase configurations.

Genetic control of Eucalyptus urophylla and E. grandis resistance to canker caused by Chrysoporthe cubensis

Chrysophorte cubensis induced canker occurs in nearly all tropical and subtropical regions where eucalypts are planted, causing losses in both wood quality and volume productivity, especially so in the warmer and more humid regions of Brazil. The wide inter and intra-specific genetic variability of resistance to canker among Eucalyptus species facilitates the selection of resistant plants. In this study, we evaluated resistance to this pathogen in five Eucalyptus grandis (G) and 15 E. urophylla (U) trees, as well as in 495 individuals from 27 progenies derived from crosses between the trees. In the field, six-months-old test seedlings were inoculated with C. cubensis. Lesion length in the xylem and bark was measured eight months later. The results demonstrated that xylem lesions could preferentially be used for the selection of resistant clones. Eight trees (7 U and 1 G) were susceptible, and the remainder (8 U and 4 G) resistant. Individual narrow and broad sense heritability estimates were 17 and 81 percent, respectively, thereby suggesting that canker resistance is quantitative and highly dependent on dominance and epistasis.

Rhizobacterial promotion of eucalypt rooting and growth

A total of 107 rhizobacterial isolates, obtained from the rhizosphere of eucalypt clones were tested as rooting inducers of cuttings and mini-cuttings planted in substrate composed of carbonized rice husk and vermiculite (1:1). Cuttings and mini-cuttings were planted in conical plastic tubes containing treated and untreated (control) substrate and kept under intermittent mist irrigation at 26-28°C. After 35 days, rooting percentage and dry root matter of cuttings were evaluated. Ten isolates capable of providing gains of up to 110 percent in root formation and up to 250 percent in root biomass over non-inoculated control cuttings were selected. Gains in rooting varied according to clone and isolate tested. The greatest gains were obtained for the mini-cuttings exhibiting the lowest rooting efficiency. Among the ten isolates tested, only 3918 (code R98) and MF4 (code R87), produced 3-indole-acetic acid in vitro, at concentrations of 0.7 and 0.67 µg ml-1, respectively. Significant increases in rooting and root dry matter of cuttings grown on rhizobacteria-inoculated substrate were found when compared to untreated or indole-butyric acid (IBA) treated mini-cuttings.

Neste trabalho, testaram-se 107 rizobactérias, isoladas da rizosfera de mudas de clones de eucalipto, quanto ao seu potencial como promotoras de enraizamento de estacas e miniestacas de eucalipto, em substrato à base de casca de arroz carbonizada e vermiculita (1:1). Estacas e miniestacas foram plantadas em tubetes cônicos contendo substrato tratado e não tratado (testemunha) e foram mantidas sob nebulização intermitente de água a 26-28°C. Aos 35 dias, avaliou-se a porcentagem média de estacas enraizadas e a massa seca do sistema radicular. Dez isolados destacaram-se como indutores de enraizamento e crescimento, propiciando ganhos de até 110 por cento e de 250 por cento, respectivamente. Esses isolados também foram eficientes no enraizamento de miniestacas, cujos ganhos variaram de acordo com o clone e isolado testado. Os maiores incrementos obtidos no enraizamento de estacas foram superiores aos observados para miniestacas. Em geral, quanto menor o índice de enraizamento do clone, maior foi o ganho médio obtido com a inoculação. Apenas os isolados 3918 (código R98) e MF4 (código R87) foram capazes de produzir ácido indol-acético (AIA) in vitro, em quantidades equivalentes a 0,7 e 0,67 µg/ml de suspensão, respectivamente. Quando comparados ao tratamento de miniestacas em ácido indol butírico (AIB), estes isolados promoveram incrementos significativos na porcentagem de enraizamento e na massa seca do sistema radicular de miniestacas.