Esterase isozymes patterns of grape vine (Vitis vinifera L) are altered in response to fungicide exposure / Padrões de isozimas Esterases de videira (Vitis vinifera L) alterados em resposta à exposição a fungicidas

Current analysis characterizes the effect of different fungicides often applied for pest control on α−and ß-esterase patterns of four economically important table-wine grape cultivars (Italia, Rubi, Benitaka and Brasil) of Vitis vinifera. The α- and ß-esterase patterns in bud leaves of the cultivars were assessed by native PAGE analysis. Cabrio Top® compound inhibited EST-2, EST-5, EST-6, EST-7, EST-8, EST-9 and EST-10 carboxylesterases, whereas EST-4, EST-11, EST-12, EST-13, EST-14 acetylesterases and EST-16 carboxylesterase were detected as weakly stained bands. Carboxylesterases and acetylesterases were also detected as weakly stained bands when exposed to fungicides Orthocide 500®, Positron Duo® and Folicur PM®. No changes in α- and ß-esterase patterns were reported when the vines were exposed to the fungicides Rovral SC®, Kumulus DF®, Curzate M®, Score® or Cuprogarb 500®. The evidence of functional changes in carboxylesterase and acetylesterase levels in current study is a warning to grape producers on the dangers inherent in the indiscriminate use of potent and modern fungicides extensively used in agriculture. The inhibition effect of fungicides on esterase isozyme molecules seems to be independent of the fungicide chemical.

O presente estudo caracterizou o efeito de diferentes fungicidas comumente aplicados como medidas de controle de pragas sobre padrões de α- e ß-esterases de quatro importantes cultivares de uva de mesa (Itália, Rubi, Benitaka e Brasil) de Vitis vinifera. Os padrões de α- e ß-esterases de brotos foliares das cultivares foram avaliados por PAGE. O composto Cabrio Top® inibiu as carboxilesterases EST-2, EST-5, EST-6, EST-7, EST-8, EST-9 e EST-10, enquanto as acetilesterases EST-4, EST-11, EST-12, EST-13, EST-14 e a carboxilesterase EST-16 foram detectadas como bandas fracamente coradas. As carboxilesterases e acetilesterases também foram detectadas como bandas fracamente coradas quando expostas aos fungicidas Orthocide 500®, Positron Duo® e Folicur PM®. Não foram observadas alterações nos padrões de α- e ß-esterases quando as videiras foram expostas aos fungicidas Rovral SC®, Kumulus DF®, Curzate M®, Score® ou Cuprogarb 500®. A evidência de alterações em nível funcional em carboxilesterases e acetilesterases, apresentada neste estudo, pode servir como um alerta aos produtores de uva dos perigos inerentes ao uso indiscriminado de fungicidas potentes e modernos amplamente utilizados hoje na agricultura. O efeito dos fungicidas sobre as enzimas esterases parece ser independente do grupo químico ao qual pertence o fungicida.

A collection of popcorn as a reservoir of genes for the generation of lineages.

In the present study, we analyze the genetic structure and diversity among accessions of popcorn obtained from the CIMMYT International Maize and Wheat Improvement Center that represent the diversity available for current use by breeding programs. The main objectives were to identify SSR loci that were the best indicators of genetic diversity, to measure the genetic diversity within popcorn genotypes, and to analyze the genetic structure of the promising populations destined for use in breeding programs. The mean gene diversity of all SSR loci was 0.6352. An extremely high population differentiation level was detected (F(st) = 0.3152) with F(st) for each locus ranging from 0.1125 (Umc1229) to 0.4870 (Umc1755). Analyzing the genetic structure of eight popcorn accessions was especially important for identifying both SSR loci with high levels of heterozygosity and genotypes showing high heterozygosity (BOYA462 and ARZM13 050). This analysis should be the medium and long-term selection goal for the generation of inbred lines and the future production of new cultivars. Plant accessions ARZM05 083, ARZM13 050, and URUG298 may also be useful varieties that exhibit important agronomic characteristics that can be used through crosses to broaden the genetic basis of popcorn.

Esterase polymorphism and the analysis of genetic diversity and structure in cactus populations descended from Cereus peruvianus plants regenerated in vitro.

The genetic structure of Cereus peruvianus populations descended from cultivated plants (F(1) populations) and from plants regenerated in vitro (R(1) populations) was analyzed using α- and ß-esterase isozymes in native PAGE. The estimated proportion of polymorphic loci was higher (50%) in the R(1) populations than the F(1) populations (42.85%). The mean observed (0.5599) and expected (0.5620) heterozygosity in R(1) descendents was also higher than the rates in F(1) descendents (H (o) = 0.4142; H (e) = 0.4977). A low level of population differentiation was detected in R(1) descendents (F (st) = 0.05). In contrast, population differentiation was high in F(1) descendents (0.2583). Esterase analysis using PAGE showed that artificial selection by silvicultural management provides high genetic diversity and a large genetic basis for C. peruvianus, whereas in vitro selection from callus tissue culture involves an increase of heterozygosity levels in descendents from somaclones and a low level of interpopulational divergence.

Size of AT(n) insertions in promoter region modulates Gmhsp17.6-L mRNA transcript levels.

During earlier experiments, an SSR molecular marker (176 Soy HSP) showing high correlation (70%) with resistance/susceptibility to javanese root-knot nematode Meloidogyne javanica was identified in soybean. After being sequenced, results indicated that the SSR 176 Soy HSP marker was inserted in the promoter region of Gmhsp17.6-L gene. It was also detected in this region that resistant genotypes presented insertions between AT(31) and AT(33) in size and susceptible genotypes, AT(9). Gmhsp17.6-L gene coding region presented a perfect match in amino acid sequence in all soybean genotypes. A ribonuclease protection assay showed that Gmhsp17.6-L gene mRNA transcripts were present in all genotypes. A real-time relative quantification (qPCR) indicated in the resistant individuals higher mRNA transcripts levels, which presented in the sequencing more AT(n) insertions. These results suggest that the number of AT(n) insertions inside this promoter region could modulate up or down gene levels. Those findings can lead to the possibility of manipulating, between some limits, the mRNA transcripts levels using different sizes of AT(n) insertions.

Genetic diversity of breeding popcorn lines determined by SSR markers / Diversidad genética de líneas de mejoramiento del Zea mays determinada por marcadores SSR

Information about genetic dissimilarity is very important to corroborate genealogical relationships and to predict the most heterozygotic hybrid combinations. Eight popcorn S6 lines of diverse germplasm types were evaluated using simple sequence repeats (SSR) markers. Of a total of 51 evaluated polymorphic primers, 15 were used for polymerase chain reaction (PCR) amplification. The genetic distance was estimated by Rogers’ modified distance. The different popcorn breeding programs in Brazil are possibly using highly similar base-populations. The genetic similarity of lines P1-3 and P8-1 was lowest, while P3-3 and P8-2 were genetically more similar. The cophenetic correlation showed that the Unweighted Pair-Group Method Using Arithmetic Averages (UPGMA) was reliable to discriminate the genotypes in five groups. The clusters were consistent with the estimates of genetic identity. There was a moderate coincidence degree between the groups and genealogy of lines. Higher levels of heterozygosity are expected from crosses between the group containing lines P3-3 and P7-3 with that of P1-3 and P7-4. Crosses between lines P1-3 and P8-1 are also promising.

Generalized lineal models for the analysis of binary data from propagation experiments of Brazilian orchids

This study aimed at applying the generalized linear models (GLM) for the analysis of a germination experiment of Cattleya bicolor in which the response variable was binary. The purpose of this experiment was to assess the effects of the storage temperatures and culture mediums on the seed viability. The analyses of variance was also carried out either with or without the data transformation. All the statistical approaches indicated the importance of the storage temperature on the seed viability. But, the culture media and interaction effects were significant only by the GLM. Based on the GLM, the seeds stored at 10°C increased viability, in which the coconut medium achieved the best performance. The results emphasized the importance of adopting the GLM to improve the reliability in many situations where the response variable followed a non-normal distribution.

A técnica de propagação in vitro é considerada efetiva para fins comerciais e de conservação de orquídeas. A metodologia de modelos lineares generalizados (MLG) foi usada para analisar um experimento de germinação de Cattleya bicolor. O propósito do experimento foi avaliar os efeitos da temperatura de armazenamento e dos meios de cultivo sobre a germinação, cuja resposta foi considerada binária. Análise convencional com ou sem transformação de dados foram também realizados. Todas as abordagens estatísticas indicaram a importância da temperatura sobre a viabilidade das sementes. Entretanto, os efeitos de meios de cultivo e interação foram significativos apenas para MLG. As sementes armazenadas a 10°C incrementaram sua viabilidade, onde o meio a base de coco atingiu o melhor desempenho. Os resultados enfatizam a importância de adotar MLG, para melhorar a confiabilidade em situações onde a variável resposta segue uma distribuição distinta à normal.

Heterozygosity following half-sib recurrent selection in popcorn using isoenzyme markers

Isozyme biochemical marker may be useful tool for genomic analysis of maize populations undergoing recurrent selection. Thus, isozymes markers was utilized for assess the changes in the genetic variability and distance in a Brazilian composite population of popcorn following four cycles of recurrent selection for yield. One hundred and ninety-six half-sib families were evaluated from each cycle and the ten highest-yielding families (5.2 percent) were recombined to produce the next cycle. Isozyme analysis considered 80 seedlings per cycle. Simple linear regression equations were estimated among the allele frequencies in each locus in function of the selection cycles, the genetic distances among the cycles and the average heterozygosity per locus for each cycle. Regression analysis did not reveal any common trend for changes in allele frequencies presumably due to selection. The estimates of the number of polymorphic locus, of the mean of allele per locus and the mean heterozygosity did not reveal any reduction in variability. It was concluded that four selection cycles did not cause relevant changes in the variability or genetic distance among the selection cycles of CMS-43 popcorn population. Isozymes markers analysis showed that the number of recombined half-sib families in recurrent selection was suitable.