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1.
Cell Mol Biol Lett ; 14(2): 289-304, 2009.
Article in English | MEDLINE | ID: mdl-19115051

ABSTRACT

The plant nuclear genome is largely composed of mobile DNA, which can rearrange genomes and other individual gene structure and also affect gene regulation through various promoted activities: transposition, insertion, excision, chromosome breakage, and ectopic recombination. Ty1-copia-like retrotransposon is a widespread class of transposable elements in the plant kingdom, representing a large part of the total DNA content. Here, a novel retrotransposon-like sequence was isolated and identified as the Ty1-copia-like reverse transcriptase domain (named here CLCoy1), based on the homology of known elements. Fluorescence in situ hybridization, revealed that CLCoy1 was mainly located in telomeric and sub-telomeric regions along the Citrus chromosomes. CLCoy1 composes 3.6% of the genome and, interestingly, while transposons are mostly specific to a species, this element was identified in other Citrus species such as Citrus aurantium, Fortunella margarita and Citrus paradisi, but undetected in Poncirus trifoliata. We also determined that wounding, salt and cell culture stress produced transcriptional activation of this novel retroelement in Citrus limon. The novel Ty1-copia-like element CLCoy1 may have played a major role in shaping genome structure and size during Citrus species evolution.


Subject(s)
Citrus/genetics , Retroelements , Transcription, Genetic , Amino Acid Sequence , Molecular Sequence Data , RNA-Directed DNA Polymerase/chemistry , RNA-Directed DNA Polymerase/genetics , Sequence Alignment
2.
Mol Biotechnol ; 41(2): 123-32, 2009 Feb.
Article in English | MEDLINE | ID: mdl-18855147

ABSTRACT

Little is known about pharmaceutical and personal care products pollutants (PPCPs), but there is a growing interest in how they might impact the environment and microbial communities. The widespread use of Viagra (sildenafil citrate) has attracted great attention because of the high usage rate, the unpredictable disposal and the unknown potential effects on wildlife and the environment. Until now information regarding the impact of Viagra on microbial community in water environment has not been reported. In this research, for the first time, the genetic profile of the microbial community, developing in a Viagra polluted water environment, was evaluated by means of the 16S and 18S rRNA genes, for bacteria and fungi, respectively, amplified by polymerase chain reaction (PCR) and separated using the denaturing gradient gel electrophoresis (DGGE) technique. The DGGE results revealed a complex microbial community structure with most of the population persisting throughout the experimental period. DNA sequences from bands observed in the different denaturing gradient gel electrophoresis profiles exhibited the highest degree of identity to uncultured bacteria and fungi found previously mainly in polluted environmental and treating bioreactors. Biotransformation ability of sildenafil citrate by the microbial pool was studied and the capability of these microorganisms to detoxify a polluted water ecosystem was assessed. The bacterial and fungal population was able to degrade sildenafil citrate entirely. Additionally, assays conducted on Daphnia magna, algal growth inhibition assay and cell viability determination on HepG2 human cells showed that biotransformation products obtained from the bacterial growth was not toxic. The higher removal efficiency for sildenafil citrate and the lack of toxicity by the biotransformation products obtained showed that the microbial community identified here represented a composite population that might have biotechnological relevance to retrieve sildenafil citrate contaminated sites.


Subject(s)
Bacteria/genetics , Biodegradation, Environmental , Fungi/genetics , Genetics, Population/methods , Piperazines/metabolism , Sulfones/metabolism , Analysis of Variance , Animals , Bacteria/metabolism , Biotransformation , Cell Line , Cell Survival , Daphnia/metabolism , Electrophoresis , Fungi/metabolism , Genes, Bacterial/genetics , Genes, Fungal/genetics , Humans , Phylogeny , Piperazines/toxicity , Polymerase Chain Reaction , Purines/metabolism , Purines/toxicity , RNA, Ribosomal, 16S/genetics , RNA, Ribosomal, 18S/genetics , Sildenafil Citrate , Sulfones/toxicity
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