Search

Abstract Ppmar1 and Ppmar2 are two active mariner-like elements (MLEs) cloned from moso bamboo (Phyllostachys edulis (Carrière) J. Houz) genome possessing transposases that harbour nuclear export signal (NES) domain, but not any nuclear localization signal (NLS) domain. To understand the functions of NES in transposon activity, we have conducted two experiments, fluorescence and excision frequency assays in the yeast system. For this, by site-directed mutagenesis, three NES mutants were developed from each of the MLE. In the fluorescence assay, the mutants, NES-1, 2 and 3 along with the wild types (NES-0) were fused with fluorescent proteins, enhanced yellow fluorescent protein (EYFP) and enhanced cyan fluorescent protein (ECFP) were co-transformed into yeast system. To differentiate protein localisation under the NES influence, ECFP alone was fused to wild and mutant NES domains either on N- or C-terminal and not to EYFP. Fluorescence assay revealed that blue fluorescence of ECFP was more intense than the red fluorescence of the EYFP in the yeast cell matrix. Further, ECFP had a wider localisation in the cellular matrix, but EYFP was largely located in the nucleus. The NES-1 domain was related to the comparatively high spread of ECFP, while NES-2 and NES-3 indicated a low spread, implying that NES activity on nuclear export increased when the NES is made leucine-rich, while the signalling activity was reduced when the leucine content was lowered in the NES domain. In the transposon excision assay, the mutant and wild type NES of both the Ppmar elements were integrated into an Ade2 vector, and within the Ade2 gene. Co-transformation of the vector together with non-autonomous Ppmar transposons and NES-lacking transposases was used to assess the differential excision frequencies of the mutants NES domains. In both the MLEs, NES-1 had the highest excision suppression, which was less than half of the excision frequency of the wild type. NES-2 and NES-3 elements showed, up to three times increase in transposon excision than the wild types. The results suggested that NES is an important regulator of nuclear export of transposase in Ppmar elements and the mutation of the NES domains can either increase or decrease the export signalling. We speculate that in moso bamboo, NESs regulates the transposition activity of MLEs to maintain the genome integrity.

The tomato leafminer, Tuta absoluta (Meyrick), (Lepidoptera: Gelechiidae), is a devastating pest of tomato that has recently become a major threat to tomato production in many countries in the Mediterranean Basin, Africa and Asia. Mariner-like elements (MLEs) are class-II transposons with short inverted terminal repeats (ITR) flanking a single open reading frame (ORF) coding for a transposase. In the present study, we have isolated eight full-length MLEs from the tomato leaf miner by PCR amplification, using a single primer designed from the ITR of Hyalophora cecropia L. Mariner signature sequence D,D(34)D, as well as the conserved amino acid motifs, WVPHEL and YSPDLAP(T)D, were identified. However, sequences analysis revealed defective elements with the presence of several kinds of mutations generating stop codons, frameshifts, and non-functional transposase. Based on predicted transposase sequences, T. absoluta MLEs (Tamar1) belongs to the cecropia subfamily and clustered closely with MLEs from other Lepidoptera members suggesting vertical genetic transmission from a common ancestor. The identification of mariner-like elements in this invasive species will be useful to understand the genome dynamics in Lepidoptera. [ABSTRACT FROM AUTHOR]

Genetic diversity of natural populations is useful in biogeographical studies. Here, we apply a Bayesian method based on the coalescent model to dating biogeographical events by using published DNA sequences of wild silkworms, Bombyx mandarina, and the domesticated model organisms B. mori, both of which categorized into the order of Lepidoptera, sampled from China, Korea, and Japan. The sequences consist of the BmTNML locus and the flanking intergenic regions. The BmTNML locus is composed of cecropia-type mariner-like element (MLE) with inverted terminal repeats, and three different transposable elements (TE), including L1BM, BMC1 retrotransposons, and BmamaT1, are inserted into the MLE. Based on the genealogy defined by TE insertions/deletions (indels), we estimated times to the most recent common ancestor and these indels events using the flanking, MLE, and indels sequences, respectively. These estimates by using MLE sequences strongly correlated with those by using flanking sequences, implying that cecropia-type MLEs can be used as a molecular clock. MLEs are thought to have transmitted horizontally among different species. By using a pair of published cecropia-type MLE sequences from lepidopteran insect, an emperor moth, and a coral in Ryukyu Islands, we demonstrated dating of horizontal transmission between species which are distantly related but inhabiting geographically close region. [ABSTRACT FROM AUTHOR]

Mariner-like elements (MLEs) are class II transposable elements found in almost all eukaryotic genomes including those of plants, from which hundreds of complete and partial sequences have been elucidated. We have characterized 82 amplification fragments representing MLEs derived from diverse members of the Bambusoideae subfamily. Phylogenetic analysis of MLE transposase sequences shows that MLEs are widespread, diverse and abundant in the Bambusoideae. In addition, a molecular phylogeny of the Bambusoideae subfamily was established by using the internal transcribed spacer sequences of nuclear ribosomal DNA (ITS) information. The phylogenetic tree constructed using MLE fragment sequences was incongruent with a second tree based on ITS information. These results suggest horizontal transfer between distantly related species or the existence of an ancestral MLE polymorphism followed by divergent evolution and stochastic loss. [ABSTRACT FROM AUTHOR]