ABSTRACT
Endophytic bacteria Bacillus safensis RS95 and Pseudomonas hibiscicola RS121 were evaluated for their ability to promote the growth of rice seedlings and produce indole-acetic acid (IAA) and siderophores and to solubilize phosphates. 'Guri' rice seeds were immersed in bacterial endophyte cell suspensions (separated and two-strain mixed), as well as in Escherichia coli DH5α, phosphate-buffered saline (PBS) and water treatments (negative controls). Seeds were sown on agar-water in Petri plates placed vertically at an angle of 65°. The ability of plant growth-promoting endophytic bacteria (PGPEB) to produce IAA and siderophores was determined by Salkowski colorimetric and chrome azurol S (CAS) assays, respectively. Mineral phosphate solubilization activity was calculated by inoculating the endophytes onto medium containing insoluble phosphate. PGPEB showed a positive effect on the growth of rice seedlings, causing a mean growth of shoots and primary-roots of 60 and 67%, respectively. Bacterial strains also showed positive traits for IAA and siderophore production, as well as phosphate-solubilization activity
Subject(s)
Pseudomonas , Oryza/growth & development , Bacillus , Siderophores , Endophytes , Indoleacetic Acids/analysis , PhosphatesABSTRACT
Xanthomonas oryzae pv. oryzae (Xoo) e X. oryzae pv. oryzicola (Xoc), são os agentes causais do crestamento bacteriano da folha (CBF) e da estria bacteriana da folha (EBF), respectivamente, são pragas transmitidas por sementes e sofrem restrição no comércio internacional de arroz, por estarem ausentes em muitos países e possuírem potencial de comprometer seriamente a produção de arroz. Vinte e um isolados oriundos de sementes de arroz do Uruguai e Argentina, Gram negativos, oxidase positiva, com pigmentação amarela típica do gêneroXanthomonas e positivos no ELISA para Xoo, foram caracterizados para determinar a associação destes, com X. oryzae e seus patovares. Os isolados foram caracterizados por métodos bioquímicos, biológicos e quanto à sensibilidade a antibióticos. Análises de PCR e qPCR foram realizadas para a confirmação do gênero Xanthomonase identificação de Xoo e Xoc. A caracterização de 21 isolados de Xanthomonassp., positivos no ELISA para Xoo, mostrou variabilidade entre os isolados e distinção de Xo.
Xanthomonas oryzae pv. oryzae (Xoo) and X. oryzae pv. oryzicola (Xoc), causal agents of bacterial leaf blight (BLB) and bacterial leaf streak (BLS), respectively, are pests transmitted by seeds and suffer restriction on international rice trade, because are absent in many countries and have potential to compromise seriously rice production. Twenty-one isolates from rice seeds from Uruguay and Argentina, Gram negative, oxidase positive, with typical yellow pigmentation of genus Xanthomonas and positive in ELISA for Xoo, were characterized to determine their association with X. oryzepathovars. Isolates were characterized by biochemical, biological and antibiogram methods. PCR and qPCR analyzes were performed to confirm genus Xanthomonasand identification of Xoo and Xoc. Characterization of 21 Xanthomonassp. isolates positive on ELISA to Xoo, showed variability among isolates and distinction of Xo.
ABSTRACT
A identificação de estirpes de rizóbio tem sido feita pela especificidade por hospedeiros e ensaios microbiológicos tradicionais. Por constituírem um grupo filogeneticamente heterogêneo, diferentes técnicas moleculares têm sido empregadas para auxiliar na caracterização genética e na identificação de estirpes eficientes e competitivas para a produção de inoculantes. Este trabalho teve por objetivos caracterizar a região espaçadora 16S-23S rDNA das estirpes de rizóbios utilizadas nos inoculantes comercializados no Brasil para espécies leguminosas, utilizando a técnica da PCR em combinação com a de RFLP, e avaliar a possibilidade do uso desse marcador molecular como método auxiliar para identificação das estipes. A amplificação da região espaçadora 16-23 S rDNA das estirpes de rizóbios gerou fragmentos com tamanhos que variaram entre 700pb e 1350pb. Os produtos resultantes da amplificação foram submetidos à digestão com as endonucleases. Mps I, Dde I e Hae III. Os resultados obtidos neste estudo indicam a possibilidade do uso da técnica de PCR-RFLP da região espaçadora 16S-23S rDNA como marcador molecular para a diferenciar as estirpes de rizóbios, em complemento às técnicas microbiológicas tradicionais. Contudo, este marcador não é suficientemente discriminatório para ser usado na identificação das estirpes recomendadas para a produção de inoculantes comerciais.
The identification of strains of rhizobia has been made by host specificity and regular microbiological tests. By forming a phylogenetically heterogeneous group, different molecular techniques have been employed to assist in the genetic characterization and identification of efficient and competitive strains for production of inoculants. This study aimed to characterize the spacer region 16S-23S rDNA of the strains of rhizobia used in commercial inoculants in Brazil for legume species, using PCR combined with RFLP, and assess the possibility of using this molecular marker as an auxiliary method for identification of strains. The amplification of the 16-23 S rDNA spacer region of rhizobium strains generated fragments with sizes ranging between 700 and 1350bp. Products from the amplification were subjected to digestion with Mps I, Dde I and Hae III endonucleases. The results indicated the possibility of using the technique of PCR-RFLP of 16S-23S spacer region rDNA as molecular marker to differentiate most strains tested and recommended for production of inoculants, in addition to the traditional microbiological techniques. However, this marker is not sufficiently discriminatory to be used in the identification of the strains recommended for the production of commercial inoculants.