Thierry Langin

The strange phylogenies of transposable elements: are horizontal transfers the only explanation?

Analyses of the evolution of transposable elements reveal some inconsistencies when the phylogenies of such elements are compared to conventional phylogenies of the host species. Such discrepancies are generally interpreted as resulting from occasional horizontal transfers of transposable elements across species boundaries. This phenomenon has been clearly demonstrated for only a few elements and both its frequency and the mechanism by which it occurs remain unknown. Moreover, in many cases, the hypothesis of horizontal transfer must be compared with alternative evolutionary scenarios.

Relationships Between Transposable Elements Based Upon the Integrase-Transposase Domains: Is There a Common Ancestor?

The integrase domain of RNA-mediated elements (class I) and the transposase domain of DNA-mediated transposable elements (class II) were compared. A number of elements contain the DDE signature, which plays an important role in their integration. The possible relationships betweenmariner-Tc1 andIS elements, retrotransposons, and retroviruses were analyzed from an alignment of this region. Themariner-Tc1 superfamily, and LTR retrotransposons and retroviruses were found to be monophyletic groups. However, theIS elements of bacteria were found in several groups. These results were used to propose an evolutionary history that suggests a common ancestor for some integrases and transposases.

CLNR1, the AREA/NIT2‐like global nitrogen regulator of the plant fungal pathogen Colletotrichum lindemuthianum is required for the infection cycle

Nitrogen starvation is generally assumed to be encountered by biotrophic and hemibiotrophic plant fungal pathogens at the beginning of their infection cycle. We tested whether nitrogen starvation constitutes a cue regulating genes that are required for pathogenicity of Colletotrichum lindemuthianum, a fungal pathogen of common bean. The clnr1 (C. lindemuthianumnitrogen regulator 1) gene, the areA/nit‐2 orthologue of C. lindemuthianum, was isolated. The predicted CLNR1 protein exhibits high amino acid sequence similarities with the AREA and NIT2 global fungal nitrogen regulators. Targeted clnr1– mutants are unable to use a wide array of nitrogen sources, indicating that clnr1 is the C. lindemuthianum major nitrogen regulatory gene. The clnr1– mutants are non‐pathogenic, although few anthracnose lesions seldom occur on whole plantlets. Surprisingly, cytological analysis reveals that the clnr1– mutants are not disturbed from the penetration stage until the end of the biotrophic phase, but that they are impaired during the setting up of the necrotrophic phase. Thus, through CLNR1, nitrogen starvation constitutes a cue for the regulation of genes that are compulsory for this stage of the C. lindemuthianum infection process. Additionally, clnr1– mutants complemented with the Aspergillus nidulans areA gene are fully pathogenic, indicating that areA is able to activate the C. lindemuthianum suited genes, normally under the control of clnr1.

Molecular Analysis of a Large Subtelomeric Nucleotide-Binding-Site–Leucine-Rich-Repeat Family in Two Representative Genotypes of the Major Gene Pools of Phaseolus vulgaris

In common bean, the B4 disease resistance (R) gene cluster is a complex cluster localized at the end of linkage group (LG) B4, containing at least three R specificities to the fungus Colletotrichum lindemuthianum. To investigate the evolution of this R cluster since the divergence of Andean and Mesoamerican gene pools, DNA sequences were characterized from two representative genotypes of the two major gene pools of common bean (BAT93: Mesoamerican; JaloEEP558: Andean). Sequences encoding 29 B4-CC nucleotide-binding-site–leucine-rich-repeat (B4-CNL) genes were determined—12 from JaloEEP558 and 17 from BAT93. Although sequence exchange events were identified, phylogenetic analyses revealed that they were not frequent enough to lead to homogenization of B4-CNL sequences within a haplotype. Genetic mapping based on pulsed-field gel electrophoresis separation confirmed that the B4-CNL family is a large family specific to one end of LG B4 and is present at two distinct blocks separated by 26 cM. Fluorescent in situ hybridization on meiotic pachytene chromosomes revealed that two B4-CNL blocks are located in the subtelomeric region of the short arm of chromosome 4 on both sides of a heterochromatic block (knob), suggesting that this peculiar genomic environment may favor the proliferation of a large R gene cluster.

Do the integrases of LTR-retrotransposons and class II element transposases have a common ancestor?

The integrases of retrotransposons (class I) and retroviruses and the transposases of bacterial type elements (class II) were compared. The DDE signature that is crucial for the integration of these elements is present in most of them, except for the non-LTR retrotransposons and members of the hAT and P super-families. Alignment of this region was used to infer the relationships between class II elements, retrotransposons, and retroviruses. The mariner-Tc1 and the Pogo-Fot1 super-families were found to be closely related and probably monophyletic, as were LTR retrotransposons and retroviruses. The IS elements of bacteria were clustered in several families, some of them being closely related to the transposase of the mariner-Tc1 super-family or to the LTR retrotransposon and retrovirus integrases. These results plus that of Xiong and Eickbush (1990) were used to develop an evolutionary history suggesting a common ancestral origin(s) for the integrases and transposases containing the DDE signature. The position of the telomeric elements (Het-A and TART) was assessed by comparing their gag and reverse transcriptase domains (when present) to those of group II introns and non-LTR retrotransposons. This preliminary analysis suggests that telomeric elements may be derived from non-LTR retrotransposons.

Horizontal transmission versus ancient origin:Mariner in the witness box

The transposable elementmariner has been found in many species ofDrosophilidae, several groups of Arthropods, and more recently in Platyhelminthes as well as in a phytopathogenic fungus. In the familyDrosophilidae, the distribution ofmariner among species shows many gaps, and its geographical distribution among endemic species is restricted to Asia and Africa. Amongmariner elements in species within and outside theDrosophilidae, the similarities in nucleotide sequence and the amino acid sequence of the putative transposase reveal many phylogenetic inconsistencies compared with the conventional phylogeny of the host species. This paper discusses the contrasting hypotheses of horizontal transfer versus ancestral origin proposed to explain these results.

VARIATION IN GENOME ORGANIZATION OF THE PLANT PATHOGENIC FUNGUS COLLETOTRICHUM LINDEMUTHIANUM

Abstract The genome structure of Colletotrichum lindemuthianum in a set of diverse isolates was investigated using a combination of physical and molecular approaches. Flow cytometric measurement of genome size revealed significant variation between strains, with the smallest genome representing 59% of the largest. Southern-blot profiles of a cloned fungal telomere revealed a total chromosome number varying from 9 to 12. Chromosome separations using pulsed-field gel electrophoresis (PFGE) showed that these chromosomes belong to two distinct size classes: a variable number of small (< 2.5 Mb) polymorphic chromosomes and a set of unresolved chromosomes larger than 7 Mb. Two dispersed repeat elements were shown to cluster on distinct polymorphic minichromosomes. Single-copy flanking sequences from these repeat-containing clones specifically marked distinct small chromosomes. These markers were absent in some strains, indicating that part of the observed variability in genome organization may be explained by the presence or absence, in a given strain, of dispensable genomic regions and/or chromosomes.

Isolation of the transposable element hupfer from the entomopathogenic fungus Beauveria bassiana by insertion mutagenesis of the nitrate reductase structural gene

Abstract A transposable element has been isolated from the entomopathogenic fungus Beauveria bassiana by trapping it in the nitrate reductase structural gene, which has been cloned from this species. The element had inserted in the first exon of the nia gene and appeared to have duplicated the sequence TA at the site of insertion. It was 3336 bp long with 30-bp imperfect, inverted, terminal repeats. The element, called hupfer, contained an open reading frame encoding a 321-amino acid protein similar to the IS630- or mariner-Tc1-like transposases, and a residual sequence of about 2 kb which was not significantly similar to any published sequence. There are fewer than five copies of this transposable element present per genome in the fungus.

Nonpathogenic Strains of Colletotrichum lindemuthianum Trigger Progressive Bean Defense Responses during Appressorium-Mediated Penetration

ABSTRACT The fungal bean pathogen Colletotrichum lindemuthianum differentiates appressoria in order to penetrate bean tissues. We showed that appressorium development in C. lindemuthianum can be divided into three stages, and we obtained three nonpathogenic strains, including one strain blocked at each developmental stage. H18 was blocked at the appressorium differentiation stage; i.e., no genuine appressoria were formed. H191 was blocked at the appressorium maturation stage; i.e., appressoria exhibited a pigmentation defect and developed only partial internal turgor pressure. H290 was impaired in appressorium function; i.e., appressoria failed to penetrate into bean tissues. Furthermore, these strains could be further discriminated according to the bean defense responses that they induced. Surprisingly, appressorium maturation, but not appressorium function, was sufficient to induce most plant defense responses tested (superoxide ion production and strong induction of pathogenesis-related proteins). However, appressorium function (i.e., entry into the first host cell) was necessary for avirulence-mediated recognition of the fungus.

Horizontal Transfers and the Evolution of Transposable Elements

Since their discovery by Barbara McClintock in the 1950s (see McClintock 1984 for a review), transposable elements have been detected in all organisms in which they have been studied. Moreover, they may represent from 5% to more than 10% of the genome. Due to their intrinsic capacity to be mobile or mobilisable, these elements can have an impact on the evolution of the genomes as discussed in a special issue of Genetica (vol 86, 1992; see also McDonald 1993). The genomes can be modified in different ways such as changes in the expression of the gene closed to their insertion point (Robins and Samuleson 1992; Smith and Corces 1992), chromosomal rearrangements such as inversions (Lim 1988; Lyttle and Haymer 1992;) or deletion (Sheen et al. 1993), evolution of intron (Wessler 1989; Purugganan and Wessler 1992), and the telomeric structure (Levis et al. 1993; Biessmann et al. 1992).