Plant disease resistance genes have been shown to be subject to positive selection, particularly in the leucine rich repeat (LRR) region that may determine resistance specificity. We performed a genome-wide analysis of positive selection in members of the nucleotide binding site (NBS)-LRR gene family of Arabidopsis thaliana. Analyses were possible for 103 of 163 NBS-LRR nucleotide sequences in the genome, and the analyses uncovered substantial evidence of positive selection. Sites under positive selection were detected and identified for 10 sequence groups representing 53 NBS-LRR sequences. Functionally characterized Arabidopsis resistance genes were in these 10 groups, but several groups with extensive evidence of positive selection contained no previously characterized resistance genes. Amino acid residues under positive selection were identified, and these residues were mapped onto protein secondary structure. Positively selected positions were disproportionately located in the LRR domain (P < 0.001), particularly a nine-amino acid ␤-strand submotif that is likely to be solvent exposed. However, a substantial proportion (30%) of positively selected sites were located outside LRRs, suggesting that regions other than the LRR may function in determining resistance specificity. Because of the unusual sequence variability in the LRRs of this class of proteins, secondary-structure analysis identifies LRRs that are not identified by similarity analyses alone. LRRs also contain substantial indel variation, suggesting elasticity in LRR length could also influence resistance specificity.

The high number of duplicated genes in plant genomes provides a potential template for gene conversion and unequal crossing-over. Within a gene family these two processes can render all members homogeneous or generate diversity by reassorting variants among paralogs. The latter is especially feasible in families where gene diversity confers a selective advantage and thus conversion events are likely to be retained. Consequently, the most complete record of gene conversion is expected to be most evident in gene families commonly subjected to positive selection. Here, we describe the extent and characteristics of gene conversion and unequal crossing-over in the coding and noncoding regions of nucleotide-binding site leucine-rich repeat (NBS-LRR), receptor-like kinases (RLK), and receptor-like proteins (RLP) in the plant Arabidopsis thaliana. Members of these three gene families are associated with disease resistance and their pathogen-recognition domain is a documented target of positive selection. Our bioinformatic approach to study the major family features that may influence gene conversion revealed that in these families there is a significant association between the occurrence of gene conversion and high levels of sequence similarity, close physical clustering, gene orientation, and recombination rate. We discuss these results in the context of the overlap between gene conversion and positive selection during the evolutionary expansion of the NBS-LRR, RLK, and RLP gene families.

The four DEF paralogues shaped floral diversity in orchids in a dramatic way by modularizing the floral perianth based on a complex series of sub- and neo-functionalization events. These genes may have eliminated constraints, so that different kinds of perianth organs could then evolve individually and thus often in dramatically different ways in response to selection by pollinators or by genetic drift. We therefore argue that floral diversity in orchids may be the result of an unprecedented developmental genetic predisposition that originated early in orchid evolution.

No abstract

SUMMARYIn flowering plants, class-B floral homeotic genes encode MADS-domain transcription factors, which are key in the specification of petal and stamen identity, and have two ancient clades: DEF-like and GLO-like genes. Many species have one gene of each clade, but orchids have typically four DEF-like genes, representing ancient gene clades 1, 2, 3 and 4. We tested the 'orchid code', a combinatorial genetic model suggesting that differences between the organs of the orchid perianth (outer tepals, inner lateral tepals and labellum) are generated by the combinatorial differential expression of four DEF-like genes. Our experimental test involves highly sensitive and specific measurements, with qRT-PCR of the expression of DEF-and GLO-like genes from the distantly related Vanilla planifolia and Phragmipedium longifolium, as well as from wild-type and peloric Phalaenopsis hybrid flowers. Our findings support the first 'orchid code' hypothesis, in that absence of clade-3 and -4 gene expression distinguishes the outer tepals from the inner tepals. In contrast to the original hypothesis, however, mRNA of both clade-3 and -4 genes accumulates in wild-type inner lateral tepals and the labellum, and in labellum-like inner lateral tepals of peloric flowers, albeit in different quantities. Our data suggest a revised hypothesis where high levels of clade-1 and -2, and low levels of clade-3 and -4, gene expression specify inner lateral tepals, whereas labellum development requires low levels of clade-1 and -2 expression and high levels of clade-3 and -4 expression.