Endophytic bacteria Bacillus safensis and Pseudomonas hibiscicola and their ability to increase rice seedling growth

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

The influence of ultra-high diluted compounds on the growth and the metabolites of Oryza sativa L

The ultra-high dilutions (UHDs) have been widely used in the field of human, animal and plant treatment. In the present research, the effects of the potentized ultra-high dilutions (UHDs) on physiological and biochemical variations in Oryza sativaL. (rice) were investigated. Methods: To study the effect of UHDs (Calendula officinalisCalen. and Arnica montanaArn.) on the physiological and biochemical variations of the Oryza sativaL. (rice), 28 experiments were designed and statistically analyzed using the Design Expert 7.0.1 software over the general factorial design methodology. Three qualitative factors were studied including the UHDs/placebo usage, sterile/non-sterile experimental condition and the type and timing of the UHDs usage. The validated analysis was subjected to more extended studies on the variations in physiological growth, carbohydrate, protein content, pigment production, and amino acid patterns. To evaluate the effects of UHDs on rice, a desirable response percentage was formed from a number of healthy seedling productions, and the height percentage of the aerial parts and main roots were studied. Results: The statistical analysis resulted in a prediction model which was more than 97% correlates with experimental results. The results showed that the UHDs increased the pH variations, carbohydrate, protein and pigment levels each by ~2.5, ~1.5, ~1.4, and ~1.4 folds, respectively. Also compared to placebo, the amount and proportionof amino acids has significantly varied, showing a statistical effect on the germination and seedling growth of the rice, as well as the stress conditions caused by the sterilization process, seedlings entrance into the light and their transition into hydroponic culture medium.Conclusion: The use of UHDs leads to an increase in the production of chlorophyll, as well as carbohydrate and protein content. Moreover, it causes significant variations in the amino acid profile and the production of amino acids along with the photosynthesis, germination, and metabolism processes.(AU)

Downregulation of OsAGO17 by artificial microRNA causes pollen abortion resulting in the reduction of grain yield in rice

Background: Pollen development is an important reproductive process that directly affects pollen fertility and grain yield in rice. Argonaute (AGO) proteins, the core effectors of RNA-mediated silencing, play important roles in regulating plant growth and development. However, few AGO proteins in rice were reported to be involved in pollen development. In this study, artificial microRNA technology was used to assess the function of OsAGO17 in pollen development. Results: In this study, OsAGO17, a rice-specific gene, was specifically expressed in rice pollen grains, with the highest expression in uninucleate microspores. Downregulation of OsAGO17 by artificial microRNA technology based on the endogenous osa-miRNA319a precursor was successfully achieved. It is found that downregulation of OsAGO17 could significantly affect pollen fertility and cause pollen abortion, thus suggesting that OsAGO17 functions in rice pollen development. In addition, the downregulation of OsAGO17 mainly caused a low seed-setting rate, thereby resulting in the reduction of grain yield, whereas the downregulation of OsAGO17 did not significantly affect rice vegetative growth and other agricultural traits including number of florets per panicle, number of primary branch per panicle, and 100-grain weight. Furthermore, the result of subcellular localization analysis indicated that the OsAGO17 protein was localized to both the nucleus and the cytoplasm. Conclusion: These results represent the first report of the biological function for OsAGO17 in rice and indicate that OsAGO17 may possibly play crucial regulatory roles in rice pollen development. It helps us to better understand the mechanism of pollen development in rice.

Contribution of dark septate fungi to the nutrient uptake and growth of rice plants

ABSTRACT The use of dark septate fungi (DSE) to promote plant growth can be beneficial to agriculture, and these organisms are important allies in the search for sustainable agriculture practices. This study investigates the contribution of dark septate fungi to the absorption of nutrients by rice plants and their ensuing growth. Four dark septate fungi isolates that were identified by Internal transcribed spacer phylogeny were inoculated in rice seeds (Cv. Piauí). The resulting root colonization was estimated and the kinetic parameters Vmax and Km were calculated from the nitrate contents of the nutrient solution. The macronutrient levels in the shoots, and the NO3--N, NH4+-N, free amino-N and soluble sugars in the roots, sheathes and leaves were measured. The rice roots were significantly colonized by all of the fungi, but in particular, isolate A103 increased the fresh and dry biomass of the shoots and the number of tillers per plant, amino-N, and soluble sugars as well as the N, P, K, Mg and S contents in comparison with the control treatment. When inoculated with isolates A103 and A101, the plants presented lower Km values, indicating affinity increases for NO3--N absorption. Therefore, the A103 Pleosporales fungus presented the highest potential for the promotion of rice plant growth, increasing the tillering and nutrients uptake, especially N (due to an enhanced affinity for N uptake) and P.

Polyphasic characterization of bacteria obtained from upland rice cultivated in Cerrado soil

ABSTRACT This work aimed to characterize 20 isolates obtained from upland rice plants, based on phenotypic (morphology, enzymatic activity, inorganic phosphate solubilization, carbon source use, antagonism), genotypic assays (16S rRNA sequencing) and plant growth promotion. Results showed a great morphological, metabolic and genetic variability among bacterial isolates. All isolates showed positive activity for catalase and protease enzymes and, 90% of the isolates showed positive activity for amylase, catalase and, nitrogenase. All isolates were able to metabolize sucrose and malic acid in contrast with mannitol, which was metabolized only by one isolate. For the other carbon sources, we observed a great variability in its use by the isolates. Most isolates showed antibiosis against Rhizoctonia solani (75%) and Sclerotinia sclerotiorum (55%) and, 50% of them showed antibiosis against both pathogens. Six isolates showed simultaneous ability of antibiosis, inorganic phosphate solubilization and protease activity. Based on phylogenetic analysis of the 16S rRNA gene all the isolates belong to Bacillus genus. Under greenhouse conditions, two isolates (S4 and S22) improved to about 24%, 25%, 30% and 31% the Total N, leaf area, shoot dry weight and root dry weight, respectively, of rice plants, indicating that they should be tested for this ability under field conditions.

Relationships of rice yield and quality based on genotype by trait (GT) biplot

ABSTRACT An experiment was conducted to examine the influencing characters on rice by using 64 rice genotypes, including four local landraces, four released cultivars and 56 mutant lines (M5) derived from these genotypes, with application of the genotype by trait (GT) biplot methodology. The first two principal components (PC1 and PC2) accounted for 46.6% of total variation in 64 genotypes. The polygon view of GT biplot suggested seven sections for 64 genotypes. The vertex G38 had good amounts of grain yield, panicle length, hundred grain weight, internodes length, plant height and fertility percentage. Generally based on vector view it was demonstrated that the selection of high grain yield would be performed via thousand grain weight, panicle weight and number of filled grain per panicle. These traits should be considered simultaneously as effective selection criteria evolving high yielding rice genotypes because of their large contribution to grain yield. The genotypes G2, G4 and G7 could be considered for the developing of desirable progenies in the selection strategy of rice improvement programs. This study revealed GT biplot can graphically display the interrelationships among traits. In conclusion, it is recommended the use of GGE biplot to identify superior genotypes for simultaneous improvement of several traits.

Insect diversity in organic rice fields under two management systems of levees vegetation / Diversidade de insetos em arroz irrigado orgânico sob dois manejos da vegetação das taipas

Abstract Simplified environments characterize agroecosystems, reducing the diversity of associated plants, which are not cultivated for economic purposes, causing unbalances that can promote the emergence of cultivated plants pests, as well as the reduction of their natural enemies. Management systems that increase diversity in agroecosystems can extend the action of natural enemies of pests. Studies to understand the diversity of insects associated with rice cultivation and determine their ecological guilds can provide information about the composition and structure of such ecosystems, which can be applied to integrated pest management. Therefore, the study aimed to describe and compare groups of insects in irrigated rice fields, with organic management using two different systems of levees vegetation management, and relate them to the phenological states of rice cultivation (seedling, vegetative, and reproductive). Samples were taken in a plantation located in Águas Claras district of Viamão, RS. The total area of 18 ha was divided into two. A subarea called not cut (NC), where wild vegetation of levees was maintained, and the subarea named cut (C), where monthly cuts were made to levees vegetation, from the beginning of soil preparation until the harvest. From October 2012 to March 2013 were held weekly collections in quadrats randomly located in both the rice fields and the levees. A total of 800 insects were collected, 429 in the C subarea and 371 in the NC. There were identified 97 morphospecies in the C and 108 in NC, being 54 shared between the subareas. The captured insects were grouped into guilds: saprophages (C = 38.2%; NC = 27.5%), phytophagous (C = 28.5%; NC = 33.2%), entomophagous (grouping parasitoids and predators) (C = 29.4%; NC = 35%) and finally other insects (C = 4 %; NC = 4.3%). The peak abundance of phytophagous and entomophagous was registered in the vegetative stage of rice. At the same stage the UPGMA analysis showed that similarity in species composition was greater than 90% in the groups obtained in the paddy fields of C and NC subareas. The vegetation of levees can positively influence the presence of entomophagous in the field. Although the abundance did not change clearly, the greatest diversity in the NC areas of all the groups, may contribute to the maintenance of ecological services expanding the system resilience.

Resumo Os agroecossistemas se caracterizam por ambientes simplificados, com redução da diversidade de plantas associadas, que não são as cultivadas para fins econômicos, causando desequilíbrios que podem levar ao surgimento de insetos nocivos, assim como a diminuição de seus inimigos naturais. Sistemas de manejo que priorizem o aumento da diversidade no agroecossistema podem ampliar a ação de inimigos naturais de pragas. Estudos que busquem entender a diversidade de insetos associados ao cultivo de arroz irrigado, bem como determinar as guildas ou grupos ecológicos aos quais pertencem, podem trazer informações sobre a composição e estrutura dos ecossistemas que possam ser aplicadas no manejo integrado de pragas. Neste sentido, o estudo objetivou conhecer e comparar a diversidade de insetos entre áreas de cultivo orgânico de arroz irrigado, diferenciadas pelo manejo da vegetação das taipas e relacionar com os estádios fenológicos da cultura. As amostragens foram realizadas no distrito de Águas Claras, município de Viamão, RS. A área total de 18 ha foi subdividida em duas. Numa subárea, denominada não roçada (NR) a vegetação espontânea das taipas foi mantida, na outra, roçada (R), foram feitas roçadas mensais das taipas, desde o início do preparo do solo, até a colheita. Entre outubro de 2012 a março de 2013 realizaram-se coletas semanais, em quadrats, situados aleatoriamente tanto nas quadras de arroz quanto nas taipas. Foi coletado um total de 800 insetos, 429 na R e 371 na NR. Foram identificadas 97 morfoespécies na R e 108 na NR, das quais 54 foram compartilhadas entre as subáreas. As guildas registradas foram: saprófagos (R = 38,2%; NR = 27,5%), fitófagos (R = 28,5%; NR = 33,2%), entomófagos (reunindo parasitoides e predadores) (R = 29,4%; NR = 35%) e outros (R = 4%; NR = 4,3%). O pico de abundância de fitófagos e entomófagos foi registrado na fase vegetativa do arroz. Nesta mesma fase, a análise de UPGMA apontou que a similaridade na composição de espécies foi superior a 90% nos grupos obtidos nas lavouras das subáreas R e NR. A vegetação das taipas pode influenciar positivamente a presença de insetos entomófagos no campo. Embora a abundância não tenha variado significativamente entre as áreas, a maior diversidade na área não roçada em todos os grupos, pode contribuir na manutenção de serviços ecológicos aumentando a resiliência dos sistemas.

Influence of the vegetation management of the leeves in irrigated rice organic in diversity of Hymenoptera parasitoids / Influência do manejo da vegetação das taipas em arroz orgânico irrigado na diversidade de himenópteros parasitoides

Abstract Among the natural enemies of insect pests in rice fields, parasitoids are especially notable. To better understand the space-time dynamics of these insects, the objectives of this study were to describe and compare groups of parasitoids in organic irrigated rice fields using two management approaches for levee vegetation, and to relate them to the phenological stages of rice cultivation (the seedling, vegetative, and reproductive stages). The samples were taken in a plantation located in Viamão, RS, Brazil. The total area of 18 ha was divided into two parts: a no-cut (NC) subarea in which the wild vegetation of the levees was maintained, and a cut (C) subarea in which the levee vegetation was cut monthly. In each subarea, four Malaise traps considered as pseudo-replicas were installed and remained in the field for 24 hours at each sampling location. Collections occurred twice a month from the beginning of cultivation (October 2012) until harvest (March 2013). A total of 3,184 Hymenoptera parasitoids were collected: 2,038 individuals in the NC subarea and 1,146 in the C subarea. We identified 458 morphospecies distributed in 24 families. Mymaridae was the most abundant and Eulophidae was the richest in both subareas. A total of 198 morphospecies was shared between the subareas, including Platygastridae, Eulophidae, and Mymaridae, which were the families with the highest number of shared species. The richness and abundance of parasitoids varied according to their phenological developmental stages, with peak abundance registering during the vegetative period. The Morisita index identified three groupings, indicating a similarity that was related to the three phases of rice growth and development: seedling, vegetative and post-harvest.

Resumo Entre os inimigos naturais de insetos pragas em campos de arroz, parasitoides são especialmente notáveis. Para melhor entender a dinâmica espaço-temporal destes insetos, os objetivos desse estudo foram descrever e comparar os grupos de parasitoides em campos de arroz orgânico irrigado utilizando duas abordagens de manejo da vegetação das taipas, e relacioná-los com os estádios fenológicos da cultura (plântula, vegetativo e reprodutivo). As amostragens foram realizadas em uma plantação localizada em Viamão, RS. A área total de 18 ha foi dividida em duas partes: uma subárea não roçada (NR) na qual a vegetação espontânea das taipas foi mantida, e uma roçada (R) subárea em que a vegetação das taipas foi roçada mensalmente. Em cada subárea, quatro armadilhas Malaise consideradas pseudo-réplicas foram instaladas e mantiveram-se no campo durante 24 horas, em cada local de amostragem. As coletas ocorreram duas vezes por mês, desde o início do cultivo (outubro de 2012) até a colheita (março de 2013). Um total de 3.184 himenópteros parasitoides foram coletados: 2.038 indivíduos na subárea NR e 1.146 na R. Foram identificadas 458 morfoespécies, distribuídas em 24 famílias. Mymaridae foi a mais abundante e Eulophidae a mais rica para ambas as subáreas. Um total de 198 morfoespécies foi compartilhado entre as subáreas, incluindo Platygastridae, Eulophidae e Mymaridae, que foram as famílias com o maior número de espécies compartilhadas. A riqueza e a abundância de parasitoides variou de acordo com as fases de desenvolvimento fenológico da cultura, com pico de abundância registrado no período vegetativo. O Índice de Morisita identificou três grupamentos indicando uma similaridade relacionada às fases da cultura, plântula, vegetativo e, na pós-colheita.

Avaliação dos fitorreguladores auxina e giberelina na germinação e crescimento do arroz / Assessment of auxin and gibberellin on rice germination and growth / Evaluación de fitorreguladores auxina y giberelina en la germinación y crecimiento del arroz

O arroz (Oryza sativa L.) é uma planta da família das Poaceas (gramíneas), sendo um dos cereais mais cultivados no mundo. Os fitorreguladores como auxina e giberilina podem influenciar o crescimento e desenvolvimento das plantas como o arroz. O objetivo deste estudo foi avaliar a germinação e o crescimento do arroz sob diferentes doses de fitorreguladores (auxina e giberelina). Foram conduzidos dois experimentos, o primeiro para determinar a germinação e comprimento radicular das sementes de arroz. O segundo experimento foi conduzido para determinar a altura, massa seca da parte área (MSPA) e o conteúdo nitrogênio (N) da parte aérea (NPA). Nos dois experimentos foram testadas quatro doses de auxina + giberelina: 0 (testemunha); 200; 500 e 1000 mL ha­1. O delineamento experimental foi inteiramente casualizado com cinco repetições para cada tratamento nos dois experimentos. A germinação das sementes e o comprimento radicular foram aumentados com a aplicação de auxina e giberelina. A altura e o NPA foram aumentados significantemente com a aplicação de auxina e giberelina. Concluiu-se que a aplicação de auxina e giberelina na cultura do arroz aumentaram a germinação e o comprimento radicular das sementes. Além disso, aumentou a altura e o conteúdo de nitrogênio nas plantas de arroz.

Rice (Oryza sativa L.) is one of the most widely grown cereals in the world. Phytoregulators such as auxin and gibberellin can influence both growth and development of plants such as rice. The aim of this study was to assess the germination and growth of rice under different doses of phytoregulators (auxin and gibberellin). Two experiments were performed, one to determine the germination and root length of rice seeds, and the second one to evaluated height, shoot dry mass (SDM) and shoot nitrogen content (SNC). Both experiments tested four doses of auxin + gibberellin (0 (control), 200, 500 and 1000 mL ha-1). The experimental design was completely randomized with five repetitions for both experiments. The germination and root length of rice seedlings were increased by the application of auxin and gibberellin. Rice height and SNC were significantly increased with the application of auxin and gibberellin. Thus, it can be concluded that the application of auxin and gibberellin in rice increases germination, root length, plant height and nitrogen content.

El arroz (Oryza sativa L.) es una planta de la familia Poaceas (gramíneas), siendo uno de los cereales más cultivados en el mundo. Los fitorreguladores como auxina y giberelina pueden influir en el crecimiento y desarrollo de plantas, como el arroz. El objetivo de este estudio ha sido evaluar la germinación y el crecimiento del arroz bajo diferentes dosis de fitorreguladores (auxina y giberelina). Se realizaron dos experimentos, el primero para determinar la germinación y longitud de la raíz de las semillas de arroz. Se llevó a cabo un segundo experimento para determinar la altura, masa seca de la parte aérea (MSPA) y el contenido de nitrógeno (N) de la parte aérea (NPA). En ambos experimentos, se ensayaron cuatro dosis de auxina + giberelina: 0 (testigo); 200; 500 y 1000mL ha-1. El diseño experimental fue completamente al azar con cinco repeticiones para cada tratamiento en ambos experimentos. La germinación de las semillas y la longitud de la raíz se incrementaron mediante la aplicación de auxina y giberelina. La altura y el NPA se incrementaron de manera significativa mediante la aplicación de auxina y giberelina. Se concluye que la aplicación de auxina y giberelina en el cultivo de arroz incrementa la germinación y longitud de la raíz de las semillas. Además, incrementó la altura de plantas de arroz con nitrógeno.

Detection of DNA polymerase λ activity during seed germination and enhancement after salinity stress and dehydration in the plumules of indica rice (Oryza sativa L.).

DNA polymerase λ (DNA pol λ) is the only reported X-family DNA polymerases in plants and has been shown to play a significant role in dry quiescent seeds, growth, development and nuclear DNA repair. cDNA for DNA pol λ has been reported in Arabidopsis and japonica rice cultivar and has been characterized from E. coli expressed protein, but very little is known about its activity at protein level in plants. The enzymatic activity of DNA pol λ was studied in dry, imbibed and during different germination stages of indica rice IR-8 (salt sensitive) by in-gel activity assay to determine its physiological role in important stages of growth and development. The upstream sequence was also analyzed using plantCARE database and was found to contain several cis-acting elements, including light responsive elements, dehydration responsive elements, Myb binding sites, etc. Hence, 4-day-old germinating seedlings of IR29, a salt-sensitive, but high yielding indica rice cultivar and Nonabokra, a salt-tolerant, but low yielding cultivar were treated with water (control) or 250 mM NaCl or 20% polyethyleneglycol-6000 for 4 and 8 h. The protein was analyzed by in vitro DNA pol λ activity assay, in-gel activity assay and Western blot analysis. DNA pol λ was not detected in dry seeds, but enhanced after imbibition and detectable from low level to high level during subsequent germination steps. Both salinity and dehydration stress led to the enhancement of the activity and protein level of DNA pol λ, as compared to control tissues. This is the first evidence of the salinity or dehydration stress induced enhancement of DNA pol λ activity in the plumules of rice (Oryza sativa L.) cultivars.

Crescimento de cultivares de arroz (Oryza sativa L. ) influenciado pela inoculação com bactérias promotoras de crescimento de plantas / Growth of rice cultivars (Oryza sativa L. ) as affected by inoculation with plant growth-promoting bacteria

Este trabalho objetivou avaliar o efeito da inoculação de seis estirpes de bactérias promotoras de crescimento de plantas (BPCP), Burkholderia spp. (B7 e B16), Pseudomonas spp. (P18 e P21) e Rhizobium spp. (R65 e R82) e duas misturas contendo uma estirpe de cada gênero no crescimento de seis cultivares de arroz (BRS Bonança, BRS Aimoré, BRS Talento, BRSMG Curinga, Japonês e Bolinha). As características fisiológicas das bactérias foram determinadas em ensaios in vitro. Foi conduzido um experimento sob condições de casa de vegetação onde as BPCP foram inoculadas em cultivares de arroz, cultivados em vasos contendo 1 kg de solo, em delineamento de blocos casualizados com 4 repetições. Aos 60 dias após a emergência foram avaliados parâmetros de crescimentos relacionados à parte aérea e ao sistema radicular dos cultivares de arroz. Entre as bactérias, somente duas (B7 e R65) não apresentaram capacidade de produção in vitro de auxina. Todas as estirpes apresentaram atividade de catalase e formação de película em meio NFb semi-sólido sem N ou presença de nódulo em feijoeiro, indicando capacidade de fixação biológica de nitrogênio. A inoculação das BPCP em arroz proporcionou aumentos médios de 10% em relação ao controle para a área foliar e massa seca da parte aérea e de 9% para o comprimento e massa seca de raiz. Entre os cultivares, BRS Aimoré apresentou maior interação com as estirpes estudadas e a estirpe P21 de Pseudomonas spp. proporcionou diferença significativa em relação ao controle em parâmetros relacionados à parte aérea e ao sistema radicular.

Aiming to evaluate the effect of the inoculation of six strains of plant growth-promoting bacteria (PGPBs) Burkholderia spp. (B7 and B16), Pseudomonas spp. (P18 and P21) and Rhizobium spp. (R65 and R82) and two mixtures containing one strain of each genera on the growth of six rice cultivars (BRS Bonança, BRS Aimoré, BRS Talento, BRSMG Curinga, Japonês and Bolinha). Physiological characteristics of the bacteria were determined on in vitro assays. Rice cultivars were inoculated with PGPB under greenhouse conditions and cultivated on 1 kg pots filled with soil on a randomized block design with four replicates. At 60 days after emergence growth parameters related with the shoot and root were evaluated for rice cultivars. Among the evaluated bacteria only two (B7 and R65) were not able to produce auxin under in vitro conditions. All strains showed catalase activity and film formation in semisolid medium NFb without N or nodulation in common bean, indicating the ability of biological nitrogen fixation. Inoculation of PGPBs in rice provided increases of 10% as compared to the control regarding leaf area and dry mass of shoots and 9% for length and root dry weight. Among the cultivars BRS Aimoré showed greater interaction with the strains studied and the strain P21 of Pseudomonas spp. promoted significant difference compared to the control for the parameters related with the shoot and root system.

Detection of genes providing salinity-tolerance in rice / Detecção de genes que conferem tolerância à salinidade em planta de arroz

The present study aimed to identify salinity-tolerant genes in three cultivars (BRS-7 Taim, BRS Querência and BRS Atalanta) of Oryza sativa L. ssp. indica S. Kato and in three cultivars (BRS Bojurú, IAS 12-9 Formosa and Goyakuman) of Oryza sativa L. ssp. japonica S. Kato. Ten days after emergence seedlings were transferred to a greenhouse and placed in a 15L vessel with half strength Hoagland nutrient solution, which was changed every four days, under controlled temperature and humidity. Plants were harvested 56 days after transfer. DNA extraction was carried out by CTAB method and salinity-tolerant genes SOS and CK1 were identified by in silico research. Amplification of gene sequence was performed with in silico primers. Bands were detected by agar gel electrophoresis and visualized under ultraviolet light after staining with ethidium bromide. Gene SOS1 fragments were present in all cultivars, except in BRS Atalanta, whereas CK1 gene was present in all evaluated cultivars. Results show that salinity-tolerant genes under analysis were identified in the two sub-species.

O estudo teve como objetivo identificar genes envolvidos na tolerância à salinidade em três cultivares (BRS-7 Taim, BRS Querência e BRS Atalanta) de Oryza sativa L. ssp. indica S. Kato e em três cultivares (BRS Bojurú, IAS 12-9 Formosa e Goyakuman) de Oryza sativa L. ssp. japonica S. Kato. As plântulas foram transferidas para casa de vegetação, sob temperatura e umidade relativa controladas, dez dias após a emergência e colocadas em bacia com capacidade para 15 litros, contendo solução nutritiva de Hoagland, meia força, a qual foi mudada em intervalos de quatro dias. A coleta foi aos 56 dias após a transferência. A extração de DNA foi realizada pelo método CTAB. Os genes SOS e CK1, envolvidos na tolerância à salinidade, foram identificados por meio de pesquisa in sílico . A amplificação das sequências do gene foi realizada utilizando-se primers também desenhados in sílico. As bandas foram detectadas por eletroforese em gel de agarose e visualizadas em luz ultravioleta após coloração com brometo de etídio. Fragmentos do gene SOS1 foram encontrados em todas as cultivares, exceto para BRS Atalanta. O gene CK1 esteve presente em todas as cultivares avaliadas. Os resultados mostram que os genes estudados estão presentes em ambas as subespécies.

Effects of 5-aminolevulinic acid (ALA)-containing supernatants from selected Rhodopseudomonas palustris strains on rice growth under NaCl stress, with mediating effects on chlorophyll, photosynthetic electron transport and antioxidative enzymes

Background: Rice is globally one of the most important food crops, and NaCl stress is a key factor reducing rice yield. Amelioration of NaCl stress was assessed by determining the growth of rice seedlings treated with culture supernatants containing 5-aminolevulinic acid (ALA) secreted by strains of Rhodopseudomonas palustris (TN114 and PP803) and compared to the effects of synthetic ALA (positive control) and no ALA content (negative control). Results: The relative root growth of rice seedlings was determined under NaCl stress (50 mM NaCl), after 21 d of pretreatment. Pretreatments with 1 μM commercial ALA and 10X diluted culture supernatant of strain TN114 (2.57 μM ALA) gave significantly better growth than 10X diluted PP803 supernatant (2.11 μM ALA). Rice growth measured by dry weight under NaCl stress ordered the pretreatments as: commercial ALA N TN114 N PP803 N negative control. NaCl stress strongly decreased total chlorophyll of the plants that correlated with non-photochemical quenching of fluorescence (NPQ). The salt stress also strongly increased hydrogen peroxide (H2O2) concentration in NaCl-stressed plants. The pretreatments were ordered by reduction in H2O2 content under NaCl stress as: commercial ALA N TN114 N PP803 N negative control. The ALA pretreatments incurred remarkable increases of total chlorophyll and antioxidative activities of catalase (CAT), ascorbate peroxide (APx), glutathione reductase (GR) and superoxide dismutase (SOD); under NaCl stress commercial ALA and TN114 had generally stronger effects than PP803. Conclusions: The strain TN114 has potential as a plant growth stimulating bacterium that might enhance rice growth in saline paddy fields at a lower cost than commercial ALA.

Carbohydrate content and antioxidative potential of the seed of three edible indica rice (Oryza sativa L.) cultivars.

Rice (Oryza sativa L.) grains or seeds are known to lose much of their nutrient and antioxidant contents, following polishing. The current study was undertaken to evaluate and compare the carbohydrate content and antioxidant parameters in the unpolished and polished seeds of three edible indica rice cultivars, namely Swarna (SW), the most popular indica rice cultivar in India and aromatic or scented cultivars Gobindobhog (GB) and Pusa Basmati (PB). While both the sucrose and starch content was the maximum in PB seeds (both unpolished and polished), the amylose content was the highest in SW polished seeds. SW polished seeds were superior as compared to GB and PB cultivars in terms of total antioxidant capacity, DPPH radical scavenging and Fe(II) chelation potential, as well as the highest lipoxygenase (LOX) inhibition or H2O2 scavenging potential, probably due to the maximum accumulation of total phenolics and flavonoids, the two important antioxidants. The reducing power ability was, however, identical in both SW and GB polished seeds. The PB polished seeds were more potent in superoxide and hydroxyl scavenging, whereas GB in nitric oxide (NO) scavenging. The common observation noted after polishing of seeds was the reduction in the level of carbohydrates and antioxidant potential, though the extent of reduction varied in the three cultivars. The only exception was GB, where there was no alteration in NO scavenging potential even after polishing. Our study showed the better performance of SW polished seeds with respect to higher amylose content and majority of the tested parameters governing antioxidant capacity and radical scavenging potential, thus highlighting the greater dietary significance of SW over the other two cultivars.

Biological control of rice brown spot with native isolates of three Trichoderma species

Brown spot caused by Bipolaris oryzae is an important rice disease in Southern coast of Caspian Sea, the major rice growing region in Iran. A total of 45 Trichoderma isolates were obtained from rice paddy fields in Golestan and Mazandaran provinces which belonged to Trichoderma harzianum, T. virens and T. atroviride species. Initially, they were screened against B. oryzae by antagonism tests including dual culture, volatile and nonvolatile metabolites and hyperparasitism. Results showed that Trichoderma isolates can significantly inhibit mycelium growth of pathogen in vitro by producing volatile and nonvolatile metabolites Light microscopic observations showed no evidence of mycoparasitic behaviour of the tested isolates of Trichoderma spp. such as coiling around the B. oryzae. According to in vitro experiments, Trichoderma isolates were selected in order to evaluate their efficacy in controlling brown spot in glasshouse using seed treatment and foliar spray methods. Concerning the glasshouse tests, two strains of T. harzianum significantly controlled the disease and one strain of T. atroviride increased the seedling growth. It is the first time that the biological control of rice brown spot and increase of seedling growth with Trichoderma species have been studied in Iran.

Perspectivas del uso de bacterias rizosféricas en el control de Pyricularia grisea (Cooke Sacc) en el cultivo del arroz (Oryza sativa L) / Prospect of the use of bacteria in the control Pyricularia grisea (Cooke Sacc) on rice (Oriza sativa L)

El arroz es fuente de alimento para una gran parte de la población mundial, con alto promedio de consumo anual. En este cultivo las enfermedades de origen microbiano constituyen uno de los factores que inciden en la obtención de bajos rendimientos y calidad de los granos. La piriculariosis o añublo del arroz, causada por Pyricularia grisea, es la enfermedad más importante en este cultivo en América Latina, ya que puede provocar hasta el 100% de reducción de los rendimientos. Como parte de la estrategia de agricultura sostenible, se trata de controlar estos patógenos y lograr altos rendimientos del cultivo mediante una combinación adecuada de fertilizantes químicos y productos biológicos. En este sentido, la aplicación de inoculantes bacterianos constituidos por bacterias promotoras del crecimiento vegetal (Plant Growth- Promoting Bacteria, PGPB, por sus siglas en inglés) ha constituido una alternativa ecológica que favorece la conservación del medioambiente y el ecosistema. Este trabajo tiene como objetivo abordar el estado actual y las perspectivas de uso de bacterias rizosféricas en el control de Pyricularia grisea en el cultivo del arroz, tratando algunos temas relevantes, como son las principales enfermedades que afectan al cultivo, los géneros de PGPB más utilizados para el control de las mismas y sus principales mecanismos de acción.

Rice is an important food supply for a large part of the world population and its consumption rates are very high. Microbial diseases are one of the main causes that provoke low yields and low-quality spotted grains. Although fungi, bacteria and viruses are mostly the responsible for these losses, fungal diseases strike more frequently. Among fungi, Pyricularia grisea, the blast fungus is responsible for up to 100% of reduction in yields, being the blast the most important rice disease in Latin America. To control this pathogen, a strategy of sustainable agriculture might be developed, combining accurately chemical and biological products. PGPB based bioproducts have been considered as an eco-friendly alternative, which favours environment preservation. This work was aimed to approach the current status and outlook of the use of rhizobacteria in the biocontrol of Pyricularia grisea on rice. Main diseases attacking rice, most beneficial PGPB and its mechanisms of action will be discussed too in this review.

Characterization of seeds of selected wild species of rice (Oryza) stored under high temperature and humidity conditions.

Wild progenitors of rice (Oryza) are an invaluable resource for restoring genetic diversity and incorporating useful traits back into cultivars. Studies were conducted to characterize the biochemical changes, including SDS-PAGE banding pattern of storage proteins in seeds of six wild species (Oryza alta, O. grandiglumis, O. meridionalis, O. nivara, O. officinalis and O. rhizomatis) of rice stored under high temperature (45oC) and humidity (~100%) for 15 days, which facilitated accelerated deterioration. Under the treated conditions, seeds of different wild rice species showed decrease in per cent germination and concentrations of protein and starch, but increase in conductivity of leachate and content of sugar. The SDS-PAGE analysis of seed proteins showed that not only the total number of bands, but also their intensity in terms of thickness differed for each species under storage. The total number of bands ranged from 11 to 22, but none of the species showed all the bands. Similarity index for protein bands between the control and treated seeds was observed to be least in O. rhizomatis and O. alta, while the indices were 0.7 and 0.625 for O. officinalis and O. nivara, respectively. This study clearly showed that seed deterioration led to distinctive biochemical changes, including the presence or absence as well as altered levels of intensity of proteins. Hence, SDS-PAGE protein banding pattern can be used effectively to characterize deterioration of seeds of different wild species of rice.

Producción y formulación de prototipos de un biofertilizante a partir de bacterias nativas asociadas al cultivo de arroz (Oryza sativa) / Producing and formulating a prototype biofertiliser from a native bacterial strain associated with rice (Oryza sativa)

Los biofertilizantes son productos con base en microorganismos que están involucrados en los procesos nutritivos de las plantas. Además de los microorganismos, es necesario mejorar las condiciones de formulación de los productos para mantener la viabilidad y estabilidad en almacenamiento y campo. El objetivo de este trabajo fue evaluar formulaciones, las cuales se realizaron mezclando el ingrediente activo (bacteria fijadora de nitrógeno) en diferentes relaciones con sustancias húmicas, Polietilenglicol, Carbopol® y quelatos. Los prototipos que mantuvieron la viabilidad durante un periodo de aproximadamente tres meses se evaluaron en cultivos de arroz. Los formulados se evaluaron teniendo en cuenta la dosis aplicada (1 y 2 L/ha); el resultado de uno de los prototipos (M= 8500 kg/ha) mostró un efecto benéfico sobre la producción superando a los testigos (7625 kg/ha). Los resultados de este trabajo, además de contribuir con el aumento de la estabilidad en almacenamiento de algunos prototipos, permitieron mostrar buenos resultados en campo (producción), ratificando la importancia que tienen estos microorganismos en los agroecosistemas

Endophytic colonization of rice (Oryza sativa L.) by the diazotrophic bacterium Burkholderia kururiensis and its ability to enhance plant growth

Burkholderia kururiensis is a diazotrophic bacterium originally isolated from a polluted aquifer environment and presents a high level of similarity with the rice endophyte "B. brasilensis" species. This work assessed the ability of B. kururiensis to endophytically colonize rice plantlets by monitoring different tissues of root-inoculated plants for the presence of bacterial growth in different media, electron microscopy and by 16S rDNA analysis. Observations of roots, stems and leaves of inoculated rice plantlets by electron microscopy revealed B. kururiensis colonization predominantly on root hair zones, demonstrating endophytic colonization primarily through the endodermis, followed by spreading into xylem vessels, a possible pathway leading to aerial parts. Although indifferent for the bacterial growth itself, addition of a nitrogen source was a limiting factor for endophytic colonization. As endophytic colonization was directly associated to an enhanced plant development, production of phytohormone auxin/indole-3-acetic acid by B. kururiensis was assayed with transgenic rice plantlets containing an auxin-responsive reporter (DR5-GUS). Our findings suggest the ability of auxin production by plant-associated B. kururiensis which may have a stimulatory effect on plant development, as evidenced by activation of DR5-GUS. We hereby demonstrate, for the first time, the ability of B. kururiensis to endophytically colonize rice, promoting both plant growth and rice grain yield.

Burkholderia kururiensis é uma bactéria diazotrófica, originalmente isolada de um ambiente aquático poluído e apresenta alto nível de similaridade com a espécie endofítica "B. brasilensis" encontrada na planta de arroz. Este artigo demonstrou a habilidade de B. kururiensis colonizar endofiticamente plântulas de arroz, após esta bactéria ter sido inoculada na raiz das plantas. Esta capacidade foi confirmada pelo crescimento bacteriano em diferentes tecidos da planta, por microscopia eletrônica e pela análise do 16S rADN. Observação por microscopia eletrônica das raízes, caule e folhas das plântulas de arroz inoculadas, revelou predominância da colonização de B. kururiensis na zona pilífera da raiz, demonstrando que a colonização endofítica inicia-se na endoderme, espalha-se pelo xilema, sendo esta a possível via para a bactéria alcançar as partes aéreas. A adição de uma fonte de nitrogênio, embora não tenha influenciado no crescimento bacteriano, foi um fator limitante para a colonização endofítica. Como a colonização endofítica mostrou-se diretamente associada ao aumento no desenvolvimento da planta, a produção do fitohormônio auxina/ácido 3-indolacético pela B. kururiensis foi verificada utilizando uma plântula de arroz transgênica, contendo o repórter responsivo para auxina (DR5-GUS). Nossos resultados sugerem que a produção de auxina pela B. kururiensis é responsável pelo estímulo no desenvolvimento da planta verificado pela ativação do DR5-GUS. Neste trabalho demonstramos, pela primeira vez, a habilidade de B. kururiensis colonizar endofiticamente a planta de arroz, promovendo tanto o aumento no crescimento da planta como a produção de sementes de arroz.

Incidence of keratinophilic fungi in waterlogged condition of paddy soil.

Prevalence of keratinophilic fungi was observed in paddy field soil during different stages of cultivation viz., transplanting, tillering, milking and maturation. Out of total 76 soil samples, 65 soil samples were found to be positive for the keratinophilic fungi. Fourteen species belonging to a single genus Chrysosporium were isolated through out the cropping season. C. keratinophilum (17.1%) followed by C. tropicum (13.15%) were found to be the most dominating geophilic species. The highest percent distribution of keratinophilic fungi appeared during the milking stage (100%) of paddy cultivation, followed by the maturation stage (89.47%).