Pattern analysis in branching and axillary flowering sequences

In the architectural approach to the study of plants, a major issue is to analyse branching and axillary flowering patterns. Due to the structured expression of the branching process and the noisy character of the observed patterns, we propose an analysis framework which is both structural and probabilistic. Data take the form of sequences which naturally represent the underlying structural information of branching and axillary flowering patterns and allow the application of a large number of methods ranging from exploratory analysis to stochastic modeling. The primary aim of the proposed analysis methods is to reveal patterns not directly apparent in the data, and thus to deepen our biological understanding of the underlying mechanisms that control the branching and the axillary flowering of plants over time and space. The proposed approach is illustrated using a set of examples corresponding to different plant species and different biological or agronomic objectives.     

SATCHMO: sequence alignment and tree construction using hidden Markov models

MOTIVATION:Aligning multiple proteins based on sequence information alone is challenging if sequence identity is low or there is a significant degree of structural divergence. We present a novel algorithm (SATCHMO) that is designed to address this challenge. SATCHMO simultaneously constructs a tree and a set of multiple sequence alignments, one for each internal node of the tree. The alignment at a given node contains all sequences within its sub-tree, and predicts which positions in those sequences are alignable and which are not. Aligned regions therefore typically get shorter on a path from a leaf to the root as sequences diverge in structure. Current methods either regard all positions as alignable (e.g. ClustalW), or align only those positions believed to be homologous across all sequences (e.g. profile HMM methods); by contrast SATCHMO makes different predictions of alignable regions in different subgroups. SATCHMO generates profile hidden Markov models at each node; these are used to determine branching order, to align sequences and to predict structurally alignable regions. RESULTS: In experiments on the BAliBASE benchmark alignment database, SATCHMO is shown to perform comparably to ClustalW and the UCSC SAM HMM software. Results using SATCHMO to identify protein domains are demonstrated on potassium channels, with implications for the mechanism by which tumor necrosis factor alpha affects potassium current. AVAILABILITY: The software is available for download from http://www.drive5.com/lobster/index.htm     

LEAFY, TERMINAL FLOWER1 and AGAMOUS are functionally conserved but do not regulate terminal flowering and floral determinacy in Impatiens balsamina

In Impatiens balsamina a lack of commitment of the meristem during floral development leads to the continuous requirement for a leaf-derived floral signal. In the absence of this signal the meristem reverts to leaf production. Current models for Arabidopsis state that LEAFY (LFY) is central to the integration of floral signals and regulates flowering partly via interactions with TERMINAL FLOWER1 (TFL1) and AGAMOUS (AG). Here we describe Impatiens homologues of LFY, TFL1 and AG (IbLFY, IbTFL1 and IbAG) that are highly conserved at a sequence level and demonstrate homologous functions when expressed ectopically in transgenic Arabidopsis. We relate the expression patterns of IbTFL1 and IbAG to the control of terminal flowering and floral determinacy in Impatiens. IbTFL1 is involved in controlling the phase of the axillary meristems and is expressed in axillary shoots and axillary meristems which produce inflorescences, but not in axillary flowers. It is not involved in maintaining the terminal meristem in either an inflorescence or indeterminate state. Terminal flowering in Impatiens appears therefore to be controlled by a pathway that uses a different integration system than that regulating the development of axillary flowers and branches. The pattern of ovule production in Impatiens requires the meristem to be maintained after the production of carpels. Consistent with this morphological feature IbAG appears to specify stamen and carpel identity, but is not sufficient to specify meristem determinacy in Impatiens.     

Modelling branching patterns on 1-year-old trunks of six apple cultivars

The structure resulting from branching on 1-year-old apple tree trunks was analysed in a set of apple cultivars with diverse branching and fruiting habits. Four different lateral types borne on successive nodes were observed when vegetative and flowering fates, as well as sylleptic and proleptic branching, were taken into account. The location and grouping of lateral types along the trunk were analysed for all cultivars, but are detailed for one cultivar only. This cultivar showed a succession of zones, each zone being characterized by its composition of lateral types. Statistical models-hidden semi-Markov chains-were built to take this structure into account and to characterize the cultivar's specific branching pattern. The models showed that most of the branching zones had a similar location in the different cultivars, even though zone composition and zone length differed among cultivars. On a more detailed scale, the nodes bearing a lateral, regardless of its type, were frequently followed by latent buds. The validity of the models and their biological interpretation are discussed with respect to parent shoot dynamics, hormonal gradients and competition between neighbouring buds.     

Grouping of amino acids and recognition of protein structurally conserved regions by reduced alphabets of amino acids

Sequence alignment is a common method for finding protein structurally conserved/similar regions. However, sequence alignment is often not accurate if sequence identities between to-be-aligned sequences are less than 30%. This is because that for these sequences, different residues may play similar structural roles and they are incorrectly aligned during the sequence alignment using substitution matrix consisting of 20 types of residues. Based on the similarity of physicochemical features, residues can be clustered into a few groups. Using such simplified alphabets, the complexity of protein sequences is reduced and at the same time the key information encoded in the sequences remains. As a result, the accuracy of sequence alignment might be improved if the residues are properly clustered. Here, by using a database of aligned protein structures (DAPS), a new clustering method based on the substitution scores is proposed for the grouping of residues, and substitution matrices of residues at different levels of simplification are constructed. The validity of the reduced alphabets is confirmed by relative entropy analysis. The reduced alphabets are applied to recognition of protein structurally conserved/similar regions by sequence alignment. The results indicate that the accuracy or efficiency of sequence alignment can be improved with the optimal reduced alphabet with N around 9. Supported by the National Natural Science Foundation of China (Grant Nos. 90403120, 10474041 and 10021001) and the Nonlinear Project (973) of the NSM     

Grouping of amino acids and recognition of protein structurally conserved regions by reduced alphabets of amino acids

Sequence alignment is a common method for finding protein structurally conserved/similar regions. However, sequence alignment is often not accurate if sequence identities between to-be-aligned sequences are less than 30%. This is because that for these sequences, different residues may play similar structural roles and they are incorrectly aligned during the sequence alignment using substitution matrix consisting of 20 types of residues. Based on the similarity of physicochemical features, residues can be clustered into a few groups. Using such simplified alphabets, the complexity of protein sequences is reduced and at the same time the key information encoded in the sequences remains. As a result, the accuracy of sequence alignment might be improved if the residues are properly clustered. Here, by using a database of aligned protein structures (DAPS), a new clustering method based on the substitution scores is proposed for the grouping of residues, and substitution matrices of residues at different levels of simplification are constructed. The validity of the reduced alphabets is confirmed by relative entropy analysis. The reduced alphabets are applied to recognition of protein structurally conserved/similar regions by sequence alignment. The results indicate that the accuracy or efficiency of sequence alignment can be improved with the optimal reduced alphabet with N around 9.     

CLAGen: a tool for clustering and annotating gene sequences using a suffix tree algorithm

Most multiple gene sequence alignment methods rely on conventions regarding the score of a multiple alignment in pairwise fashion. Therefore, as the number of sequences increases, the runtime of sequencing expands exponentially. In order to solve the problem, this paper presents a multiple sequence alignment method using a linear-time suffix tree algorithm to cluster similar sequences at one time without pairwise alignment. After searching for common subsequences, cross-matching common subsequences were generated, and sometimes inexact matching was found. So, a procedure aimed at masking the inexact cross-matching pairs was suggested here. In addition, BLAST was combined with a clustering tool in order to annotate the clusters generated by suffix tree clustering. The proposed method for clustering and annotating genes consists of the following steps: (1) construction of a suffix tree; (2) searching and overlapping common subsequences; (3) grouping subsequence pairs; (4) masking cross-matching pairs; (5) clustering gene sequences; (6) annotating gene clusters by the BLAST search. The performance of the proposed system, CLAGen, was successfully evaluated with 42 gene sequences in a TCA cycle (a citrate cycle) of bacteria. The system generated 11 clusters and found the longest subsequences of each cluster, which are biologically significant.     

The Flowering Unit in the Synflorescences of Amaranthaceae

The structure of the synflorescence and the flowering units in Amaranthaceae are characterized. The synflorescence is polytelic. In the flowering unit we recognize the main florescence and the enrichment zone. The florescences may consist of: (1) Fully developed partial florescences bearing three or more flowers; (2) Partial florescences reduced to one or a few fertile flowers having prophylls with more or less modified axillary productions; or (3) No partial florescences but solitary flowers having prophylls with no axillary productions. We described the flowering unit in species with florescences bearing a solitary flower and the flowering unit in species with florescences bearing partial florescences. Hypothesized developmental processes are described, with a view to finding relationships among different models characterized in the family as well as defining characters for cladistic studies, which may be useful to depict all the variations observed.     

Efficient methods for multiple sequence alignment with guaranteed error bounds

Multiple string (sequence) alignment is a difficult and important problem in computational biology, where it is central in two related tasks: finding highly conserved subregions or embedded patterns of a set of biological sequences (strings of DNA, RNA or amino acids), and inferring the evolutionary history of a set of taxa from their associated biological sequences. Several precise measures have been proposed for evaluating the goodness of a multiple alignment, but no efficient methods are known which compute the optimal alignment for any of these measures in any but small cases. In this paper, we consider two previously proposed measures, and given two computationaly efficient multiple alignment methods (one for each measure) whose deviation from the optimal value isguaranteed to be less than a factor of two. This is the novel feature of these methods, but the methods have additional virtues as well. For both methods, the guaranteed bounds are much smaller than two when the number of strings is small (1.33 for three strings of any length); for one of the methods we give a related randomized method which is much faster and which gives, with high probability, multiple alignments with fairly small error bounds; and for the other measure, the method given yields a non-obviouslower bound on the value of the optimal alignment.     

Sample size for a phylogenetic inference.

The objective of this work is to describe sample-size calculations for the inference of a nonzero central branch length in an unrooted four-species phylogeny. Attention is restricted to independent binary characters, such as might be obtained from an alignment of the purine-pyrimidine sequences of a nucleic acid molecule. A statistical test based on a multinomial model for character-state configurations is described. The importance of including invariable sites in models for sequence change is demonstrated, and their effect on sample size is quantified. The methods are applied to a four-species alignment of small-subunit rRNA sequences derived from two archaebacteria, a eubacteria and a eukaryote. We conclude that the information in these sequences is not sufficient to resolve the branching order of this tree. Estimates of the number of aligned nucleotide positions required to provide a reasonably powerful test are given.     

Micrografting techniques for testing long-distance signalling in Arabidopsis

Grafting in species other than Arabidopsis has generated persuasive evidence for long-distance signals involved in many plant processes, including regulation of flowering time and shoot branching. Hitherto, such approaches in Arabidopsis have been hampered by the lack of suitable grafting techniques. Here, a range of micrografting methods for young Arabidopsis seedlings are described. The simplest configuration was a single-hypocotyl graft, constructed with or without a supporting collar, allowing tests of root-shoot communication. More complex two-shoot grafts were also constructed, enabling tests of shoot-shoot communication. Integrity of grafts and absence of adventitious roots on scions were assessed using plants constitutively expressing a GUS gene as one graft partner. Using the max1 (more axillary growth) and max3 increased branching mutants, it was shown that a wild-type (WT) rootstock was able to inhibit rosette branching of mutant shoots. In two-shoot grafts with max1 and WT shoots on a max1 rootstock, the mutant shoot branched profusely, but the WT one did not. In two-shoot grafts with max1 and WT shoots on a WT rootstock, neither shoot exhibited increased branching. The results mirror those previously demonstrated in equivalent grafting experiments with the ramosus mutants in pea, and are consistent with the concept that a branching signal is capable of moving from root to shoot, but not from shoot to shoot. These grafting procedures will be valuable for revealing genes associated with many other long-distance signalling pathways, including flowering, systemic resistance and abiotic stress responses.     

3MATRIX and 3MOTIF: a protein structure visualization system for conserved sequence motifs

Computational methods such as sequence alignment and motif construction are useful in grouping related proteins into families, as well as helping to annotate new proteins of unknown function. These methods identify conserved amino acids in protein sequences, but cannot determine the specific functional or structural roles of conserved amino acids without additional study. In this work, we present 3MATRIX (http://3matrix.stanford.edu) and 3MOTIF (http://3motif.stanford.edu), a web-based sequence motif visualization system that displays sequence motif information in its appropriate three-dimensional (3D) context. This system is flexible in that users can enter sequences, keywords, structures or sequence motifs to generate visualizations. In 3MOTIF, users can search using discrete sequence motifs such as PROSITE patterns, eMOTIFs, or any other regular expression-like motif. Similarly, 3MATRIX accepts an eMATRIX position-specific scoring matrix, or will convert a multiple sequence alignment block into an eMATRIX for visualization. Each query motif is used to search the protein structure database for matches, in which the motif is then visually highlighted in three dimensions. Important properties of motifs such as sequence conservation and solvent accessible surface area are also displayed in the visualizations, using carefully chosen color shading schemes.     

Improving the performance of protein threading using insertion/deletion frequency arrays

As a protein evolves, not every part of the amino acid sequence has an equal probability of being deleted or for allowing insertions, because not every amino acid plays an equally important role in maintaining the protein structure. However, the most prevalent models in fold recognition methods treat every amino acid deletion and insertion as equally probable events. We have analyzed the alignment patterns for homologous and analogous sequences to determine patterns of insertion and deletion, and used that information to determine the statistics of insertions and deletions for different amino acids of a target sequence. We define these patterns as insertion/deletion (indel) frequency arrays (IFAs). By applying IFAs to the protein threading problem, we have been able to improve the alignment accuracy, especially for proteins with low sequence identity. We have also demonstrated that the application of this information can lead to an improvement in fold recognition.