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The obligate mutualism and exquisite specificity of many plant-pollinator interactions lead to the expectation that flower phenotypes (e.g., corolla tube length) and corresponding pollinator traits (e.g., hawkmoth proboscis length) are congruent as a result of coevolution by natural selection. However, the effect of variation in flower morphology on the fitness of plants and their pollinators has not been quantified systematically. In this study, we employed the theoretical morphospace paradigm using a combination of 3D printing, electronic sensing, and machine vision technologies to determine the influence of two flower morphological features (corolla curvature and nectary diameter) on the fitness of both parties: the artificial flower and its hawkmoth pollinator. Contrary to the expectation that the same flower morphology maximizes the fitness of both plant and pollinator, we found that the two parties have divergent optima for corolla curvature, with non-overlapping fitness peaks in flower morphospace. The divergent fitness optima between plants and pollinators could lead to evolutionary diversification in both groups., Competing Interests: The authors have declared that no competing interests exist.

Conspicuous differences in floral morphology are partly responsible for reproductive isolation between two sympatric species of monkeyflower because of their effect on visitation of the flowers by different pollinators. Mimulus lewisii flowers are visited primarily by bumblebees, whereas M. cardinalis flowers are visited mostly by hummingbirds. The genetic control of 12 morphological differences between the flowers of M. lewisii and M. cardinalis was explored in a large linkage mapping population of F2 plants n = 465 to provide an accurate estimate of the number and magnitude of effect of quantitative trait loci (QTLs) governing each character. Between one and six QTLs were identified for each trait. Most (9/12) traits appear to be controlled in part by at least one major QTL explaining >/=25% of the total phenotypic variance. This implies that either single genes of individually large effect or linked clusters of genes with a large cumulative effect can play a role in the evolution of reproductive isolation and speciation.

Poplar trees have at least two different chitinase genes, win6 and win8, which are systemically wound-inducible and belong to multigene families [Proc Natl Acad Sci USA 86: 7895-7899]. On one genomic clone that we have partially sequenced, there are three win6 genes which are transcriptionally oriented in the same direction. Between two of the win6 genes is a gene that we have designated chitinase X (chiX), which appears to be a pseudogene belonging to a multigene family distinct from win6 and win8. The win6 and chiX genes we have sequenced contain two AT-rich introns that correspond in location to those in a basic chitinase gene from tobacco. The predicted Win6 proteins have a putative signal peptide, a cysteine-rich 'hevein' domain, a hinge region, and a catalytic domain as described in Shinshi et al. [Plant Mol Biol 14: 357-368]. The predicted Win8 protein, by contrast, completely lacks a hinge region. Both Win6 and Win8 are expected to be highly acidic (with a calculated net charge of -15 to -17), whereas ChiX proteins are likely to be basic. Based on an inferred phylogeny, the catalytic domain of ChiX is more closely related to the basic chitinases of herbaceous plants than are either Win6 or Win8.

When the lower leaves of hybrid poplar trees are mechanically wounded, several novel mRNAs accumulate in the unwounded upper leaves (Parsons TJ, Bradshaw HD, Gordon MP: Systemic accumulation of specific mRNAs in response to wounding in poplar trees, Proc Natl Acad Sci USA, in press). A partial cDNA clone corresponding to a transcript from the wound-responsive gene designated win 3 (wound-inducible) has been cloned by differential hybridization to 32P-labelled cDNA from the leaves of wounded trees. Northern blots show a large accumulation of win 3 transcripts in the unwounded leaves of wounded trees. Southern blot analysis of poplar DNA suggests that win 3 is a member of a multigene family. The nucleotide sequences of several win 3 cDNA clones have been determined, indicating that at least three win 3 gene family members are transcribed. A genomic clone of a win 3 gene family member has been isolated and a 1.5 kb Hind III fragment containing the predicted protein-coding and 5' upstream regions has been sequenced. The putative win 3 gene product is similar to the major soluble proteins of sweet potato tubers, sporamin A and sporamin B. Both Win3 and the sporamins share significant amino acid sequence identity with Kunitz-type trypsin inhibitors from legume seeds. The Kunitz family of proteinase inhibitors thus joints three other proteinase inhibitor families which are systemically responsive to wounding.

We have designed and constructed an electroporation device capable of efficient transfer of DNA into both plant cell protoplasts and cultured murine lymphocytes. The electroporator design allows various combinations of voltage and capacitance to be used to optimize the electric pulse. Switching of large voltages and currents is accomplished with a silicon-controlled rectifier, yielding excellent reproducibility and long component life. A safety switch is provided to permit complete discharge of the device. Conditions suitable for high levels of transient expression and high frequencies of stable transformation for both plant and animal cell systems have been found.

cDNAs for several transcripts that accumulate systemically in response to mechanical wounding in a hybrid poplar have been cloned. The corresponding mRNAs become abundant in the unwounded upper leaves of poplar trees whose lower leaves have been damaged. Two of the cDNA clones have been characterized by Northern and Southern blotting, and their nucleotide sequences have been determined. These clones, designated win6 (wound-inducible) and win8, are members of multigene families encoding proteins with a high degree of similarity to chitinases from bean, tobacco, and barley. This initial demonstration of a systemic response to wounding in trees provides an approach to study defense mechanisms in woody plants at the biochemical, physiological, and ecological levels.