Construcción de una filogenia molecular para las especies de los géneros Klebsiella y Raoultella basada en los genes ARNr 16S y ARN polimerasa subunidad

Las bacterias de los géneros Raoultella y Klebsiella son patógenos oportunistas para las cuales no existe un sistema uniforme de clasificación taxonómica internacional. En el presente estudio se propone una filogenia molecular basada en el gen ribosomal 16S (ADNr 16S) y el gen codificante de la subunidad de la ARN polimerasa (rpoB) de los géneros Klebsiella y Raoultella con el fin de establecer relaciones evolutivas entre dichos géneros. Los resultados evidencian una agrupación acorde con la taxonomía y las propiedades bioquímicas características, reportadas en el Genbank. Se estableció una bifurcación en los árboles, lo cual confirma la separación de los géneros Klebsiella y Raoultella. Adicionalmente, se confirmó el carácter polifilético de K. aerogenes por el gen ADNr 16S y la agrupación de R. terrigena y K. oxytoca de acuerdo con el gen rpoB. La comparación entre los árboles obtenidos permitió determinar relaciones evolutivas entre las especies, a partir de los genes evaluados, lo cual refleja cambios aparentes a nivel taxonómico y corrobora la importancia del análisis a nivel de multilocus. Este tipo de estudios permite monitorear la estabilidad de los genotipos microbianos sobre la escala temporal y espacial, mejorar la precisión de las anotaciones taxonómicas (mejor descripción de taxones o subdivisiones genéticas) y evaluar la diversidad genética y adaptabilidad en términos de virulencia o resistenciaa drogas.

Correlação entre medidas paramétricas e não-paramétricas de estabilidade em soja

Foram avaliadas a adaptabilidade e a estabilidade de genótipos de soja (Glycine max L.) segundo a metodologia clássica de Eberhart e Russell e a estabilidade dos mesmos genótipos pela metodologia não-paramétrica de Huhn. Os experimentos foram conduzidos no delineamento em blocos casualizados, com três repetições e com 30 tratamentos (genótipos de soja), durante três anos consecutivos. As parcelas experimentais foram constituídas por quatro linhas de cultivo, espaçadas de 0,50 m e com densidade de 25 plantas por metro linear. Como área útil, foram tomadas as linhas centrais, eliminando-se 0,5 m de cada extremidade. A comparação entre as metodologias foi efetuada considerando-se o caráter produção de grãos. Verificou-se correlação de posição significativa dos postos dos genótipos, entre o desvio da regressão e as duas medidas não-paramétricas de estabilidade, porém o mesmo não foi observado entre o coeficiente de regressão e as medidas não-paramétricas (Si(1) e Si(2)). As medidas Si(1) e Si(2) mostraram-se quase que perfeitamente correlacionadas.

Temporal stability for unpredictable annual climatic variability for Hevea genotype selection

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Genome stability pathways in head and neck cancers

Genomic instability underlies the transformation of host cells toward malignancy, promotes development of invasion and metastasis and shapes the response of established cancer to treatment. In this review, we discuss recent advances in our understanding of genomic stability in squamous cell carcinoma of the head and neck (HNSCC), with an emphasis on DNA repair pathways. HNSCC is characterized by distinct profiles in genome stability between similarly staged cancers that are reflected in risk, treatment response and outcomes. Defective DNA repair generates chromosomal derangement that can cause subsequent alterations in gene expression, and is a hallmark of progression toward carcinoma. Variable functionality of an increasing spectrum of repair gene polymorphisms is associated with increased cancer risk, while aetiological factors such as human papillomavirus, tobacco and alcohol induce significantly different behaviour in induced malignancy, underpinned by differences in genomic stability. Targeted inhibition of signalling receptors has proven to be a clinically-validated therapy, and protein expression of other DNA repair and signalling molecules associated with cancer behaviour could potentially provide a more refined clinical model for prognosis and treatment prediction. Development and expansion of current genomic stability models is furthering our understanding of HNSCC pathophysiology and uncovering new, promising treatment strategies. © 2013 Glenn Jenkins et al.

Comparative analysis of Tritrichomonas foetus (Riedmuller, 1928) cat genotype, T. foetus (Riedmuller, 1928) cattle genotype and Tritrichomonas suis (Davaine, 1875) at 10 DNA loci

The parasitic protists in the genus Tritrichomonas cause significant disease in domestic cattle and cats. To assess the genetic diversity of feline and bovine isolates of Tritrichomonas foetus (Riedmuller, 1928) Wenrich and Emmerson, 1933, we used 10 different genetic regions, namely the protein coding genes of cysteine proteases 1,2 and 4-9 (CP1, 2, 4-9) involved in the pathogenesis of the disease caused by the parasite. The cytosolic malate dehydrogenase 1 (MDH1) and internal transcribed spacer region 2 of the rDNA unit (ITS2) were included as additional markers. The gene sequences were compared with those of Tritrichomonas suis (Davaine. 1875) Morgan and Hawkins, 1948 and Tritrichomonas mobilensis Culberson et al., 1986. The study revealed 100% identity for all 10 genes among all feline isolates (=T. foetus cat genotype), 100% identity among all bovine isolates (=T. foetus cattle genotype) and a genetic distinctness of 1% between the cat and cattle genotypes of T. foetus. The cattle genotype of T. foetus was 100% identical to T. suis at nine loci (CP1, 2,4-8, ITS2, MDH1). At CP9, three out of four T. suis isolates were identical to the T. foetus cattle genotype, while the T. suis isolate SUI-H3B sequence contained a single unique nucleotide substitution. Tritrichomonas mobilensis was 0.4% and 0.7% distinct from the cat and cattle genotypes of T. foetus, respectively. The genetic differences resulted in amino acid changes in the CP genes, most pronouncedly in CP2, potentially providing a platform for elucidation of genotype-specific host-pathogen interactions of T. foetus. On the basis of this data we judge T. suis and T. foetus to be subjective synonyms. For the first time, on objective nomenclatural grounds, the authority of T. suis is given to Davaine, 1875, rather than the commonly cited Gruby and Delafond, 1843. To maintain prevailing usage of T. foetus, we are suppressing the senior synomym T. suis Davaine, 1875 according to Article 23.9, because it has never been used as a valid name after 1899 and T. foetus is widely discussed as the cause of bovine trichomonosis. Thus bovine, feline and porcine isolates should all be given the name T. foetus. This promotes the stability of T. foetus for the veterinary and economically significant venereal parasite causing bovine trichomonosis. (C) 2012 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

Micropropagation of Begonia and a study of genome stability in Begonia rex

The development of procedures and media for the micropropagation of B. rex are described. Media for the production of plantlets from a number of other Begonia hybrids are also provided. Growth analysis data is given for plants produced in vivo from leaf cuttings and in vitro from mature leaf petioles and immature leaves derived from singly and multiply recycled axenic plantlets. No significant difference was found in phenotype or quantitative vegetative characters for any of the populations assessed. The results presented from studies on the development of broad spectrum media for the propagation of a number of B. rex cultivars using axenic leaf explants on factorial combinations of hormones illustrate the major influence played by the genotype on explant response in vitro and suggest media on which a range of B. rex cultivars may be propagated. Procedures for in vitro irradiation and colchicine treatments to destabilize the B. rex genome have also been described. Variants produced from these treatments indicate the utility of in vitro procedures for the expression of induced somatic variation. Colour variants produced from irradiation treatment have been cultured and prove stable. Polyploids produced as variants from irradiation treatment have been subcultured but prove unstable. Media for the induction and proliferation of callus are outlined. The influence of callus subculture and aging on the stability of the B. rex genome is assessed by chromosomal analysis of cells, in vitro and in regenerants. The B. rex genome is destabilized in callus culture but attenuation of variation occurs on regeneration. Diploid cell lines are maintained in callus subcultures and supplementation of regenerative media with high cytokinin concentrations, casein hydrolysate or adenine failed to produce variants. Callus aging however resulted in the production of polyploids. The presence and expression of pre-existing somatic variation in B. rex pith and root tissue is assessed and polyploids have been produced from pith tissues cultured in vitro. The stability of the B. rex genome and the application of tissue culture to micropropagation and breeding of B. rex are discussed.

Tumor Risks and Genotype-Phenotype-Proteotype Analysis in 358 Patients With Germline Mutations in SDHB and SDHD

Succinate dehydrogenase B (SDHB) and D (SDHD) subunit gene mutations predispose to adrenal and extraadrenal pheochromocytomas, head and neck paragangliomas (HNPGL), and other tumor types. We report tumor risks in 358 patients with SDHB (n = 295) and SDHD (n = 63) mutations. Risks of HNPGL and pheochromocytoma in SDHB mutation carriers were 29% and 52%, respectively, at age 60 years and 71% and 29%, respectively, in SDHD mutation carriers. Risks of malignant pheochromocytoma and renal tumors (14% at age 70 years) were higher in SDHB mutation carriers; 55 different mutations (including a novel recurrent exon 1 deletion) were identified. No clear genotype-phenotype correlations were detected for SDHB mutations. However, SDHD mutations predicted to result in loss of expression or a truncated or unstable protein were associated with a significantly increased risk of pheochromocytoma compared to missense mutations that were not predicted to impair protein stability (most such cases had the common p.Pro81Leu mutation). Analysis of the largest cohort of SDHB/D mutation carriers has enhanced estimates of penetrance and tumor risk and supports in silicon protein structure prediction analysis for functional assessment of mutations. The differing effect of the SDHD p.Pro81Leu on HNPGL and pheochromocytoma, risks suggests differing mechanisms of tumorigenesis in SDH-associated HNPGL and pheochromocytoma. Hum Mutat 31:41-51, 2010. (C) 2009 Wiley-Liss, Inc.

Prediction of Hevea progeny performance in the presence of genotype-environment interaction

Twenty two open-pollinated Hevea progenies from different parental clones of the Asian origin were tested at five sites in the Northwestern São Paulo State Brazil to investigate the progeny girth growth, rubber yield, bark thickness and plant height. Except for the rubber yield, the analysis of variance indicated highly significant (p<0.01) genotype x environment interaction and heterogeneity of regressions among the progenies. However, the regression stability analysis identified only a few interacting progenies which had regression coefficients significantly different from the expected value of one. The linear regressions of the progeny mean performance at each test on an environmental index (mean of all the progenies in each test) showed the general stability and adaptability of most selected Hevea progenies over the test environments. The few progenies which were responsive and high yielding on different test sites could be used to maximize the rubber cultivars productivity and to obtain the best use of the genetically improved stock under different environmental conditions.

A Bayesian approach for assessing the stability of genotypes

Several statistical models can be used for assessing genotype X environment interaction (GEI) and studying genotypic stability. The objectives of this research were to show how (i) to use Bayesian methodology for computing Shukla's phenotypic stability variance and (ii) to incorporate prior information on the parameters for better estimation. Potato [Solanum tuberosum subsp. andigenum (Juz. & Bukasov) Hawkes], wheat (Triticum aestivum L.), and maize (Zea mays L.) multi environment trials (MET) were used for illustrating the application of the Bayes paradigm. The potato trial included 15 genotypes, but prior information for just three genotypes was used. The wheat trial used prior information on all 10 genotypes included in the trial, whereas for the maize trial, noninformative priors for the nine genotypes was used. Concerning the posterior distribution of the genotypic means, the maize MET with 20 sites gave less disperse posterior distributions of the genotypic means than did the posterior distribution of the genotypic means of the other METs, which included fewer environments. The Bayesian approach allows use of other statistical strategies such as the normal truncated distribution (used in this study). When analyzing grain yield, a lower bound of zero and an upper bound set by the researcher's experience can be used. The Bayesian paradigm offers plant breeders the possibility of computing the probability of a genotype being the best performer. The results of this study show that although some genotypes may have a very low probability of being the best in all sites, they have a relatively good chance of being among the five highest yielding genotypes.

Rubber tree early selection for yield stability in time and among locations

Rubber production in the rubber tree [Hevea brasiliensis (Willd. ex Adr. de Juss.) Muell. Arg.] can be expressed differently in different environments. Thus the objective of the present study was to select productive progenies, stable and responsive in time and among locations. Thirty progenies were assessed by early yield tests at three ages and in three locations. A randomized block design was used with three replications and ten plants per plot, in 3 × 3 m spacing. The procedure of the mixed linear Reml/Blup model-restricted maximum likelihood/best non-biased linear prediction was used in the genetic statistical analyses. In all the individual analyses, the values observed for the progeny average heritability (ĥpa 2) were greater than those of the additive effect based on single individuals (ĥa 2) and within plot additive (ĥad 2). In the joint analyses in time, there was genotype × test interaction in the three locations. When 20 % of the best progenies were selected the predicted genetic gains were: Colina GG = 24.63 %, Selvíria GG = 13.63 %, and Votuporanga GG = 25.39 %. Two progenies were among the best in the analyses in the time and between locations. In the joint analysis among locations there was only genotype × location interaction in the first early test. In this test, selecting 20 %, the general predicted genetic gain was GG = 25.10 %. Identifying progenies with high and stable yield over time and among locations contributes to the efficiency of the genetic breeding program. The relative performance of the progenies varies depending of the age of early selection test. © 2013 Springer Science+Business Media Dordrecht.

Genetic parameters and simultaneous selection for root yield, adaptability and stability of cassava genotypes

O objetivo deste trabalho foi estimar parâmetros genéticos e avaliar a seleção simultânea quanto à produtividade de raízes e à adaptabilidade e estabilidade de genótipos de mandioca. Os efeitos dos genótipos foram considerados como fixos e aleatórios, e a metodologia de modelos mistos (REML/Blup) foi utilizada para estimar os parâmetros genéticos e a média harmônica do desempenho relativo dos valores genotípicos (MHPRVG), para seleção simultânea. Dez genótipos foram avaliados em delineamento de blocos ao acaso, com quatro repetições. O experimento foi realizado nos municípios de Altamira, Santarém e Santa Luzia do Pará, PA, nos anos agrícolas de 2009/2010, 2010/2011 e 2011/2012. As raízes foram colhidas 12 meses após o plantio, em todos os locais testados. A produtividade de raízes apresentou baixo coeficiente de variação genotípica (4,25%) e herdabilidade de parcelas individuais no sentido amplo (0,0424), o que resultou em baixo ganho genético. Em razão da baixa correlação genotípica (0,15), a classificação dos genótipos quanto à produtividade de raízes variou de acordo com o ambiente. Os genótipos CPATU 060, CPATU 229 e CPATU 404 destacaram-se quanto à produtividade, adaptabilidade e estabilidade.

Productivity, stability and adaptability in open pollination progenies of Eucalyptus urophylla ST Blake

Prediction of genetic gains within breeding programs is not always compatible with those observed in practice. One reason for this inconsistency is the lack of knowledge of genotype-environment interaction (GxE). The aim of this study was to estimate genetic variation, evaluate the GxE, investigate the genetic correlation between pairs of environments and for the set, and to study the productivity, stability and adaptability at 2 years of age for diameter at breast height (DBH) in five progenies trials of Eucalyptus urophylla, used in a randomized complete block design, with the number of progenies ranging from 138 to 167, four to eight blocks and five to six plants per plot. Estimates of variance components and genetic parameters were obtained using the REML/BLUP method. For analysis of productivity, stability and adaptability, the HMRPGV method was used. The highest DBH growth was observed in Anhembi (10.52 cm) and Uberaba (10.20 cm). Estimates considered high were obtained for the coefficient of individual additive genetic variation (>13.3%) and average heritability among progenies (>0.40), indicating the possibility of obtaining genetic gains by selection among progenies. The coefficient of determination of the GxE was 1.7%, a fact that led to a high value of genotypic correlation between the performance of the progenies and environments (78.1%), indicating that the interaction is simple. The first six progenies showed a coincidence of 100% in the order of stability (HMGV), adaptability (RPGV) and productivity (HMRPGV), being 13% higher than the overall mean of five experiments (9.21 cm). When ordering the progenies, the selection of the 20 best in growth led to an increase in gain ranging of from 10.4 to 70%. Anhembi is the ideal place to have a breeding population which will be good in the other places as well.

Comparative analysis of Tritrichomonas foetus (Riedmüller, 1928) cat genotype, T. foetus (Riedmüller, 1928) cattle genotype and Tritrichomonas suis (Davaine, 1875) at 10 DNA loci.

The parasitic protists in the genus Tritrichomonas cause significant disease in domestic cattle and cats. To assess the genetic diversity of feline and bovine isolates of Tritrichomonas foetus (Riedmüller, 1928) Wenrich and Emmerson, 1933, we used 10 different genetic regions, namely the protein coding genes of cysteine proteases 1, 2 and 4-9 (CP1, 2, 4-9) involved in the pathogenesis of the disease caused by the parasite. The cytosolic malate dehydrogenase 1 (MDH1) and internal transcribed spacer region 2 of the rDNA unit (ITS2) were included as additional markers. The gene sequences were compared with those of Tritrichomonas suis (Davaine, 1875) Morgan and Hawkins, 1948 and Tritrichomonas mobilensisCulberson et al., 1986. The study revealed 100% identity for all 10 genes among all feline isolates (=T. foetus cat genotype), 100% identity among all bovine isolates (=T. foetus cattle genotype) and a genetic distinctness of 1% between the cat and cattle genotypes of T. foetus. The cattle genotype of T. foetus was 100% identical to T. suis at nine loci (CP1, 2, 4-8, ITS2, MDH1). At CP9, three out of four T. suis isolates were identical to the T. foetus cattle genotype, while the T. suis isolate SUI-H3B sequence contained a single unique nucleotide substitution. Tritrichomonas mobilensis was 0.4% and 0.7% distinct from the cat and cattle genotypes of T. foetus, respectively. The genetic differences resulted in amino acid changes in the CP genes, most pronouncedly in CP2, potentially providing a platform for elucidation of genotype-specific host-pathogen interactions of T. foetus. On the basis of this data we judge T. suis and T. foetus to be subjective synonyms. For the first time, on objective nomenclatural grounds, the authority of T. suis is given to Davaine, 1875, rather than the commonly cited Gruby and Delafond, 1843. To maintain prevailing usage of T. foetus, we are suppressing the senior synomym T. suisDavaine, 1875 according to Article 23.9, because it has never been used as a valid name after 1899 and T. foetus is widely discussed as the cause of bovine trichomonosis. Thus bovine, feline and porcine isolates should all be given the name T. foetus. This promotes the stability of T. foetus for the veterinary and economically significant venereal parasite causing bovine trichomonosis.

Functional and physiological consequences of genetic variation at phosphoglucose isomerase: Heat shock protein expression is related to enzyme genotype in a montane beetle

Allele frequency variation at the phosphoglucose isomerase (PGI) locus in Californian populations of the beetle Chrysomela aeneicollis suggests that PGI may be undergoing natural selection. We quantified (i) apparent Michaelis-Menten constant (Km) of fructose 6-phosphate at different temperatures and (ii) thermal stability for three common PGI genotypes (1–1, 1–4, and 4–4). We also measured air temperature (Ta) and beetle body temperature (Tb) in three montane drainages in the Sierra Nevada, California. Finally, we measured 70-kDa heat shock protein (Hsp70) expression in field-collected and laboratory-acclimated beetles. We found that PGI allele 1 predominated in the northernmost drainage, Rock Creek (RC), which was also significantly cooler than the southernmost drainage, Big Pine Creek (BPC), where PGI allele 4 predominated. Allele frequencies and air temperatures were intermediate in the middle drainage, Bishop Creek (BC). Differences among genotypes in Km (1–1 > 1–4 > 4–4) and thermal stability (4–4 > 1–4 > 1–1) followed a pattern consistent with temperature adaptation. In nature, Tb was closely related to Ta. Hsp70 expression in adult beetles decreased with elevation and differed among drainages (BPC > BC > RC). After laboratory acclimation (8 days, 20°C day, 4°C night) and heat shock (4 h, 28–36°C), Hsp70 expression was greater for RC than BPC beetles. In RC, field-collected beetles homozygous for PGI 1–1 had higher Hsp70 levels than heterozygotes or a 4–4 homozygote. These results reveal functional and physiological differences among PGI genotypes, which suggest that montane populations of this beetle are locally adapted to temperature.

The evaluation of the spatial and temporal stability of sugarcane farm performance based on yield and commercial cane sugar

In broader catchment scale investigations, there is a need to understand and ultimately exploit the spatial variation of agricultural crops for an improved economic return. In many instances, this spatial variation is temporally unstable and may be different for various crop attributes and crop species. In the Australian sugar industry, the opportunity arose to evaluate the performance of 231 farms in the Tully Mill area in far north Queensland using production information on cane yield (t/ha) and CCS ( a fresh weight measure of sucrose content in the cane) accumulated over a 12-year period. Such an arrangement of data can be expressed as a 3-way array where a farm x attribute x year matrix can be evaluated and interactions considered. Two multivariate techniques, the 3-way mixture method of clustering and the 3-mode principal component analysis, were employed to identify meaningful relationships between farms that performed similarly for both cane yield and CCS. In this context, farm has a spatial component and the aim of this analysis was to determine if systematic patterns in farm performance expressed by cane yield and CCS persisted over time. There was no spatial relationship between cane yield and CCS. However, the analysis revealed that the relationship between farms was remarkably stable from one year to the next for both attributes and there was some spatial aggregation of farm performance in parts of the mill area. This finding is important, since temporally consistent spatial variation may be exploited to improve regional production. Alternatively, the putative causes of the spatial variation may be explored to enhance the understanding of sugarcane production in the wet tropics of Australia.