A molecular rheostat adjusts auxin flux to promote root protophloem differentiation.

Auxin influences plant development through several distinct concentration-dependent effects 1 . In the Arabidopsis root tip, polar auxin transport by PIN-FORMED (PIN) proteins creates a local auxin accumulation that is required for the maintenance of the stem-cell niche2-4. Proximally, stem-cell daughter cells divide repeatedly before they eventually differentiate. This developmental gradient is accompanied by a gradual decrease in auxin levels as cells divide, and subsequently by a gradual increase as the cells differentiate5,6. However, the timing of differentiation is not uniform across cell files. For instance, developing protophloem sieve elements (PPSEs) differentiate as neighbouring cells still divide. Here we show that PPSE differentiation involves local steepening of the post-meristematic auxin gradient. BREVIS RADIX (BRX) and PROTEIN KINASE ASSOCIATED WITH BRX (PAX) are interacting plasma-membrane-associated, polarly localized proteins that co-localize with PIN proteins at the rootward end of developing PPSEs. Both brx and pax mutants display impaired PPSE differentiation. Similar to other AGC-family kinases, PAX activates PIN-mediated auxin efflux, whereas BRX strongly dampens this stimulation. Efficient BRX plasma-membrane localization depends on PAX, but auxin negatively regulates BRX plasma-membrane association and promotes PAX activity. Thus, our data support a model in which BRX and PAX are elements of a molecular rheostat that modulates auxin flux through developing PPSEs, thereby timing PPSE differentiation.

Fast and accurate phylogenetic reconstruction from high-resolution whole-genome data and a novel robustness estimator.

The rapid accumulation of whole-genome data has renewed interest in the study of genomic rearrangements. Comparative genomics, evolutionary biology, and cancer research all require models and algorithms to elucidate the mechanisms, history, and consequences of these rearrangements. However, even simple models lead to NP-hard problems, particularly in the area of phylogenetic analysis. Current approaches are limited to small collections of genomes and low-resolution data (typically a few hundred syntenic blocks). Moreover, whereas phylogenetic analyses from sequence data are deemed incomplete unless bootstrapping scores (a measure of confidence) are given for each tree edge, no equivalent to bootstrapping exists for rearrangement-based phylogenetic analysis. We describe a fast and accurate algorithm for rearrangement analysis that scales up, in both time and accuracy, to modern high-resolution genomic data. We also describe a novel approach to estimate the robustness of results-an equivalent to the bootstrapping analysis used in sequence-based phylogenetic reconstruction. We present the results of extensive testing on both simulated and real data showing that our algorithm returns very accurate results, while scaling linearly with the size of the genomes and cubically with their number. We also present extensive experimental results showing that our approach to robustness testing provides excellent estimates of confidence, which, moreover, can be tuned to trade off thresholds between false positives and false negatives. Together, these two novel approaches enable us to attack heretofore intractable problems, such as phylogenetic inference for high-resolution vertebrate genomes, as we demonstrate on a set of six vertebrate genomes with 8,380 syntenic blocks. A copy of the software is available on demand.

NETGEN: generating phylogenetic networks with diploid hybrids.

SUMMARY: NetGen is an event-driven simulator that creates phylogenetic networks by extending the birth-death model to include diploid hybridizations. DNA sequences are evolved in conjunction with the topology, enabling hybridization decisions to be based on contemporary evolutionary distances. NetGen supports variable rate lineages, root sequence specification, outgroup generation and many other options. This note describes the NetGen application and proposes an extension of the Newick format to accommodate phylogenetic networks. AVAILABILITY: NetGen is written in C and is available in source form at http://www.phylo.unm.edu/~morin/.

A linear-time algorithm for computing inversion distance between signed permutations with an experimental study.

Hannenhalli and Pevzner gave the first polynomial-time algorithm for computing the inversion distance between two signed permutations, as part of the larger task of determining the shortest sequence of inversions needed to transform one permutation into the other. Their algorithm (restricted to distance calculation) proceeds in two stages: in the first stage, the overlap graph induced by the permutation is decomposed into connected components; then, in the second stage, certain graph structures (hurdles and others) are identified. Berman and Hannenhalli avoided the explicit computation of the overlap graph and gave an O(nalpha(n)) algorithm, based on a Union-Find structure, to find its connected components, where alpha is the inverse Ackerman function. Since for all practical purposes alpha(n) is a constant no larger than four, this algorithm has been the fastest practical algorithm to date. In this paper, we present a new linear-time algorithm for computing the connected components, which is more efficient than that of Berman and Hannenhalli in both theory and practice. Our algorithm uses only a stack and is very easy to implement. We give the results of computational experiments over a large range of permutation pairs produced through simulated evolution; our experiments show a speed-up by a factor of 2 to 5 in the computation of the connected components and by a factor of 1.3 to 2 in the overall distance computation.

New approaches for reconstructing phylogenies from gene order data.

We report on new techniques we have developed for reconstructing phylogenies on whole genomes. Our mathematical techniques include new polynomial-time methods for bounding the inversion length of a candidate tree and new polynomial-time methods for estimating genomic distances which greatly improve the accuracy of neighbor-joining analyses. We demonstrate the power of these techniques through an extensive performance study based on simulating genome evolution under a wide range of model conditions. Combining these new tools with standard approaches (fast reconstruction with neighbor-joining, exploration of all possible refinements of strict consensus trees, etc.) has allowed us to analyze datasets that were previously considered computationally impractical. In particular, we have conducted a complete phylogenetic analysis of a subset of the Campanulaceae family, confirming various conjectures about the relationships among members of the subset and about the principal mechanism of evolution for their chloroplast genome. We give representative results of the extensive experimentation we conducted on both real and simulated datasets in order to validate and characterize our approaches. We find that our techniques provide very accurate reconstructions of the true tree topology even when the data are generated by processes that include a significant fraction of transpositions and when the data are close to saturation.

A new implementation and detailed study of breakpoint analysis.

Phylogenies derived from gene order data may prove crucial in answering some fundamental open questions in biomolecular evolution. Yet very few techniques are available for such phylogenetic reconstructions. One method is breakpoint analysis, developed by Blanchette and Sankoff for solving the "breakpoint phylogeny." Our earlier studies confirmed the usefulness of this approach, but also found that BPAnalysis, the implementation developed by Sankoff and Blanchette, was too slow to use on all but very small datasets. We report here on a reimplementation of BPAnalysis using the principles of algorithmic engineering. Our faster (by 2 to 3 orders of magnitude) and flexible implementation allowed us to conduct studies on the characteristics of breakpoint analysis, in terms of running time, quality, and robustness, as well as to analyze datasets that had so far been considered out of reach. We report on these findings and also discuss future directions for our new implementation.

A new fast heuristic for computing the breakpoint phylogeny and experimental phylogenetic analyses of real and synthetic data.

The breakpoint phylogeny is an optimization problem proposed by Blanchette et al. for reconstructing evolutionary trees from gene order data. These same authors also developed and implemented BPAnalysis [3], a heuristic method (based upon solving many instances of the travelling salesman problem) for estimating the breakpoint phylogeny. We present a new heuristic for this purpose; although not polynomial-time, our heuristic is much faster in practice than BPAnalysis. We present and discuss the results of experimentation on synthetic datasets and on the flowering plant family Campanulaceae with three methods: our new method, BPAnalysis, and the neighbor-joining method [25] using several distance estimation techniques. Our preliminary results indicate that, on datasets with slow evolutionary rates and large numbers of genes in comparison with the number of taxa (genomes), all methods recover quite accurate reconstructions of the true evolutionary history (although BPAnalysis is too slow to be practical), but that on datasets where the rate of evolution is high relative to the number of genes, the accuracy of all three methods is poor.

First Report of Rhizoctonia solani AG-3 on Potato in Catalonia (NE Spain).

During the last 3 years, potatoes (Solanum tuberosum L.) with symptoms typical of Rhizoctonia canker and black scurf have been found in Catalonia; from 21,546 samples submitted to the official diagnostic laboratory (Laboratori de Sanitat Vegetal, DARP, Generalitat de Catalunya), 20% were infected with R. solani. Also, basidia and basidiospores of Thanatephorus cucumeris (A.B. Frank) Donk, the teleomorph of R. solani, have been observed frequently on stems near the soil surface. Previous work (1) has reported R. solani in the same geographic area, but anastomosis groups (AG) were not identified. Isolates (103) were collected from damaged roots, mature or progeny tubers, soil, debris, and weeds (roots and stem of Oxalis latifolia, Diplotaxis eurocoides, Solanum nigrum, Sorghum halepense, and Chenopodium album) in fields where potatoes were cultivated. All isolates were multinucleate, variable in number of nuclei (mean = 12, range = 7 to 19), and with morphological features typical of R. solani (3). Following the procedure of Parmeter et al. (2), all isolates (84) were identified as R. solani AG-3. Thirty-two isolates were evaluated for pathogenicity on potato plants at 20 to 25°C in the greenhouse. Germinated tubers (sprout length 3 cm) free of Rhizoctonia were placed in pea-sand mix (3:1, vol/vol; Compo Sana Universal, Barcelona, Spain) at 5-cm depth and inoculated by placing a 5-mm-diameter plug cut from 1-week-old cultures of each isolate adjacent to the sprouts. Potatoes were harvested 20 days after planting, and both emerged and nonemerged sprouts were examined for lesions and rated using a 0 to 4 scale. The AG-3 isolates caused major damage to roots and shoots (average disease rating 3.4). R. solani isolates were recovered from diseased plants and identified as AG-3. Preliminary field studies in Catalonia (Baix Llobregat and Maresme, unpublished) indicate that disease severity is not dependent on cultivar or related to yield reduction (10 to 65%), but is related to soil and seed contamination with sclerotia and mycelium. This is the first reported occurrence of R. solani AG-3 on potato in Catalonia. References: (1) Cebolla et al. Actas del III Congreso de la SECH, 1989. (2) Parmeter et al. Phytopathology 59:1270, 1969. (3) Sneh et al. Identification of Rhitzoctonia species. American Phytopathological Society, St. Paul, MN, 1991.