YMA Medium Research Articles

Caracterização de rizóbios isolados de Jacatupé cultivado em solo salino no Estado de Pernanbuco, Brasil

Pesquisas sobre a biodiversidade microbiológica de solos salinos envolvem a busca por genótipos tolerantes a esse tipo de estresse ambiental. Dados genotípicos correlacionados às características morfológicas, fisiológicas e bioquímicas de bactérias fornecem informações importantes para sua identificação e agrupamento. Este trabalho objetivou caracterizar rizóbios provenientes de solos salinos, do Agreste e Sertão de Pernambuco, utilizando jacatupé (Pachyrhizus erosus L. Urban) como planta-isca. Os testes foram efetuados em meio YMA e as características culturais observadas em 24 isolados foram as seguintes: mudança de pH, tempo de crescimento, transparência, forma, borda, produção de exopolissacarídeo das colônias e resistência à salinidade. Os testes moleculares utilizando análise por PCR (reação em cadeia da polimerase) por meio de seqüências repetitivas de DNA amplificadas com o primer BOX envolveram 13 isolados. Os resultados revelaram alta diversidade fenotípica e genotípica entre os isolados nativos. As características culturais e genéticas desses isolados foram comparados com 19 estirpes de referência. Os isolados NFB746 e NFB747 tiveram alta semelhança entre si e também com as estirpes Rhizobium sp. NGR234 (BR2406) e Mesorhizobium ciceri USDA3383 (BR521). O isolado NFB742, possivelmente, era da mesma espécie de M. ciceri (BR521). Com relação ao isolado NFB741, a semelhança com as bactérias Rhizobium tropici IIA CFN299T (BR10016) e Sinorhizobium terangue USDA4894 (BR527) foi de 87%. Os demais isolados, praticamente, formaram grupos independentes quando comparados com as estirpes de referência. Os resultados foram de grande relevância para diagnosticar novas espécies de rizóbios nativos altamente tolerantes a estresses ambientais.

Produksi Pigmen Karotenoid oleh Khamir Phaffia rhodozyma yang Diperlakukan dengan Radiasi Sinar UV

Carotenoid pigment is an essential element in aquaculture, since it gives characteristic of color on shrimp and fish. Carotenoid pigments can be produced microbiologically using Paffia rhodozyma . Genetic improvement of the yeast, one of which can be accomplished by radiation mutation, will increase the production of carotenoid pigments. The aims of this study were to mutate P. rhodozyma using UV irradiation and to figure out pigment production by the mutant strains resulting from 30 minute-irradiation. Irradiated culture was incubated in dark condition and plated onto YMA media. Grown mutant colonies were collected in order to test for their pigment production. Pigment production was measured on the basis of extinction coefficient of 1%. The results showed that mutant strain encoded with MUV-1 produced the highest pigment at 179.96 mg/g dry weight cell, higher than the wild type (63.20 mg/g dry weight cell).

The Transcriptional Response of Saccharomyces cerevisiae to Pichia membranifaciens Killer Toxin

The transcriptional response of Saccharomyces cerevisiae to Pichia membranifaciens killer toxin (PMKT) was investigated. We explored the global gene expression responses of the yeast S. cerevisiae to PMKT using DNA microarrays, real time quantitative PCR, and Northern blot. We identified 146 genes whose expression was significantly altered in response to PMKT in a non-random functional distribution. The majority of induced genes, most of them related to the high osmolarity glycerol (HOG) pathway, were core environmental stress response genes, showing that the coordinated transcriptional response to PMKT is related to changes in ionic homeostasis. Hog1p was observed to be phosphorylated in response to PMKT implicating the HOG signaling pathway. Individually deleted mutants of both up- (99) and down-regulated genes (47) were studied for altered sensitivity; it was observed that the deletion of up-regulated genes generated hypersensitivity (82%) to PMKT. Deletion of down-regulated genes generated wild-type (36%), resistant (47%), and hypersensitive (17%) phenotypes. This is the first study that shows the existence of a transcriptional response to the poisoning effects of a killer toxin.

Inhibition of crown gall formation by Agrobacterium radiobacter biovar 3 strains isolated from grapevine

Graft unions of nursery stock of grapevine (Vitis vinifera L.) collected in Japan yielded pathogenic and nonpathogenic strains of Agrobacterium. On the basis of classical diagnostic tests, a sequence analysis, and a multiplex polymerase chain reaction method previously reported, the pathogenic strain was identified as Agrobacterium tumefaciens biovar 3, whereas the nonpathogenic strains were assigned to Agrobacterium radiobacter biovar 3. Stems of tomato (Lycopersicon esculentum Mill.) seedlings were inoculated with both A. tumefaciens biovar 3 strain G-Ag-27 as a pathogen and one of the control strains isolated from grapevine or A. radiobacter biovar 2 strain K84 as competitors to assay the suppression of gall formation caused by the pathogen. In a test with a 1 : 1 pathogen/nonpathogen cell ratio, all A. radiobacter biovar 3 strains reduced gall incidence and size compared to that of the positive control inoculated only with the pathogen. Strain VAR03-1 was especially effective in reducing the incidence of gall formation on grapevine and reduced gall size by 84%–100% of those on the positive control. Many tested nonpathogenic biovar 3 strains were bacteriocinogenic, causing an inhibition zone against A. tumefaciens biovar 3 strains on YMA medium. Strain VAR03-1 was the most effective against indicator strains and appears to be a promising agent for controlling crown gall of grapevine.

Physiological Characteristics of Symbiotic Fungi Associated with the Seed Germination of Gastrodia elata

This study was carried out to investigate the possibility for seeds germination of Gastrodia elata using symbiotic fungi. Since seeds of G. elata are very small and lack an endosperm and other nutrients, their germination is difficult without requirement for external nutrients. Out of twenty six isolates collected from protocorms of G. elata and roots of native orchids inhabited in wild, two strains (H-2 and H-21) were observed to stimulate the seed germination of G. elata. The seed germination of G. elata was excellent on oak tree leaves medium. The optimal conditions for mycelial growth of symbiotic fungi were 25°C and pH 6.0, respectively. The mycelial growth of H-2 strain was excellent on YMA medium, while H-21 was poor on PDA medium. In case of carbon sources, the mycelial growth of H-2 and H-21 was good on media supplemented with glucose and dextrin, respectively. Calcium nitrate was good for mycelial growth of H-2 strain as a nitrogen sources, whereas urea was effective to H-21 strain.

Tolerância de rizóbios de diferentes procedências ao zinco, cobre e cádmio

Sessenta estirpes/isolados dos gêneros Bradyrhizobium, Rhizobium, Sinorhizobium, Mesorhizobium e Azorhizobium, procedentes de diferentes locais (Mata Atlântica, Amazônia, culturas agrícolas e experimentos com metais pesados) e de espécies hospedeiras pertencentes às subfamílias Papilionoideae, Mimosoideae e Caesalpinoideae, foram avaliadas quanto à tolerância a Zn, Cu e Cd em meio YMA modificado pela adição de tampões biológicos (HEPES e MES) e suplementados com Cu (0 a 60 mg L-1), Cd (0 a 60 mg L-1) e Zn (0 a 1.000 mg L-1). Mediante padrões de crescimento atribuídos às culturas nas diferentes concentrações dos metais, avaliaram-se as concentrações máximas toleradas e as doses tóxicas destes metais para redução de crescimento em 25% (DT25) e 50% (DT50). Não houve influência da procedência na concentração máxima de metal tolerada. A ordem de sensibilidade aos metais, considerando-se as concentrações máximas toleradas, foi Azorhizobium > Rhizobium = Mesorhizobium = Sinorhizobium > Bradyrhizobium. A DT25 e a DT50 foram úteis para diferenciarem estirpes/isolados de um mesmo gênero, que atingiram a mesma concentração máxima tolerada a Zn, Cu e Cd. A ordem de toxicidade dos metais estudados foi Cu > Cd > Zn.

Quantitative evaluation of acidity tolerance of root nodule bacteria

Quantification of acidity tolerance in the laboratory may be the first step in rhizobial strain selection for the Amazon region. The present method evaluated rhizobia in Petri dishes with YMA medium at pH 6.5 (control) and 4.5, using scores of 1.0 (sensitive, "no visible" growth) to 4.0 (tolerant, maximum growth). Growth evaluations were done at 6, 9, 12, 15 and 18 day periods. This method permits preliminary selection of root nodule bacteria from Amazonian soils with statistical precision. Among the 31 rhizobia strains initially tested, the INPA strains 048, 078, and 671 presented scores of 4.0 at both pHs after 9 days of growth. Strain analyses using a less rigorous criterion (growth scores higher than 3.0) included in this highly tolerant group the INPA strains 511, 565, 576, 632, 649, and 658, which grew on the most diluted zone (zone 4) after 9 days. Tolerant strains still must be tested for nitrogen fixation effectiveness, competitiveness for nodule sites, and soil persistence before their recommendation as inoculants.