Effects of floral diversity and pollinator behaviour on the persistence of hybrid zones between plants sharing pollinators

Background: Plant hybrid zones often display a large diversity in floral traits due to segregation and recombination occurring in the second and later hybrid generations. Such diversity can have important effects on pollinator behaviour.

Aims: We hypothesised that the number of floral types and their relative densities may influence pollinator preferences and level of flower constancy.

Methods: To test the influence of floral diversity on the number of visits to inflorescences of the most common type and on the level of flower constancy, we monitored bumblebees’ behaviour in experimental arrays of artificial inflorescences with a common (‘parental’) floral type mixed with either one or three uncommon (‘hybrid’) floral types.

Results: We found that pollinators preferentially visited the most common flower colour and showed slightly greater constancy when the number of uncommon floral types was increased.

Conclusions: We conclude that diversity in flower colour could promote moderate pollinator constancy in some types of hybrid zones. Segregation of floral phenotypes due to hybridisation may result in small levels of pollinator-mediated assortative mating and contribute, to a small extent, and most probably in combination with other factors, to the persistence of some hybrid zones.     

Induction of adventitious shoots and tetraploids in Antirrhinum majus L. by treatment of antimitotic agents in vitro without plant growth regulators

We examined the effects of five antimitotic agents using Antirrhinum majus L. ‘Maryland True Pink’ on the induction of adventitious shoots resulted in increase of frequencies of chromosome doubling without plant growth regulators. Seeds were treated in vitro with 0, 16.5, 32.9, 65.8, 131.6, or 263.2 µM oryzalin (ORY), amiprofos-methyl (APM), butamifos (BUT), or propham (IPC) or 800, 1,600, 3,200, 6,400, or 12,800 µM colchicine (COL) for 7 day. ORY, COL and APM promoted induction of adventitious shoots on the hypocotyls at maximum frequencies of 57.6% with 16.5 µM ORY, 5.6% with 800 µM COL and 88.8% with 131.6 µM APM. ORY and COL also induced adventitious shoots on the epicotyls adjacent to the cotyledons, particularly at high concentrations, with a maximum frequency of 26.0% at 12,800 µM COL. APM treatment increased frequencies of tetraploids from 0.0 to 93.1%, with a positive correlation between the frequency and concentration. By contrast, ORY and COL induced tetraploids at frequencies of 16.0 to 54.6% and 4.0 to 59.4%, respectively, with peaks at both low and high concentrations of each. Correlation analysis revealed that frequencies of adventitious shoot formation could be useful as an index for the induction of tetraploids. These results showed that three of the antimitotic agents tested induced both adventitious shoot and tetraploid without plant growth regulators, indicating that antimitotic action may play a common role in the induction of adventitious shoot.     

Patterns of floral colour neighbourhood and their effects on female reproductive success in an Antirrhinum hybrid zone

The maintenance of genetic integrity of parental populations is often explained by selection against hybrids. However, the selection agents are usually unknown. The role of environmental biotic interactions is often suspected but has rarely been demonstrated. In plants for instance, mutualism with pollinators may be involved. After verification that pollen deposition is a limiting factor for fruit set, we used an individual-based study and a representation of pollinator colour perception to test the effects of local plant density and floral colour neighbourhood on female reproductive success in an Antirrhinum hybrid zone. In addition to flower colour and density effects, the composition of the floral neighbourhood was found to influence fruit set, suggesting that most plants were usually better fertilized when similar to their neighbours. However, the plants of one particular type were sometimes favoured when very different from their neighbours. The implications for hybrid zone dynamics are discussed.     

Why do paralogs persist? Molecular evolution of CYCLOIDEA and related floral symmetry genes in Antirrhineae (Veronicaceae)

CYCLOIDEA (CYC) and DICHOTOMA (DICH) are paralogous genes that determine adaxial (dorsal) flower identity in the bilaterally symmetric flowers of Antirrhinum majus (snapdragon). We show here that the duplication leading to the existence of both CYC and DICH in Antirrhinum occurred before the radiation of the Antirrhineae (the tribe to which snapdragon belongs). We find no additional gene duplications within Antirrhineae. Using explicit codon-based models of evolution in a likelihood framework, we show that patterns of molecular evolution after the duplication that gave rise to CYC and DICH are consistent with purifying selection acting at both loci, despite their known functional redundancy in snapdragon. However, for specific gene regions, purifying selection is significantly relaxed across DICH lineages, relative to CYC lineages. In addition, we find evidence for relaxed purifying selection along the lineage leading to snapdragon in one of two putative functional domains of DICH. A model of selection accounting for the persistence of paralogous genes in the absence of diversifying selection is presented. This model takes into account differences in the degree of purifying selection acting at the two loci and is consistent with subfunctionalization models of paralogous gene evolution.     

Allozymic differentiation of the Antirrhinum graniticum and the Antirrhinum meonanthum species groups

Forty populations from eight taxa belonging to six species of Antirrhinum were studied for electrophoretic differentiation at 14 allozyme loci. All the studied species show that marker alleles and genetic distances between conspecific populations are lower than for other species of the genus. Results permit the recognition of A. onubensis and A. boissieri at specific rank. The low level of genetic distance together with morphological characters and sympatric range, support the recognition of A. ambiguum as a new subspecies of A. graniticum, as A. graniticum ssp. ambiguum Mateu & Segarra stat. nov. The results of the study support separation of the studied taxa into two groups coinciding with subsections Streptosepalum and Antirrhinum, but disagreeing with the arrangement of species into them. According to the results of the study these groups consist of A. meonanthum and A. braun-blanquetii (Streptosepalum) and A. graniticum s.l., A. onubensis and A. boissieri (Antirrhinum). The status of A. meonanthum var. rothmaleri requires further clarification.     

Interactions between gene activity and cell layers during floral development

The DEFICIENS (DEF) gene is required for establishing petal and stamen identity in Antirrhinum and is expressed in all three layers of the floral meristem in whorls 2 and 3. Expression of DEF in a subset of meristem layers gives rise to organs with characteristic shapes and cell types, reflecting altered patterns and levels of DEF gene activity. To determine how the contributions of layers and gene activity interact, we exploited a DEF allele which carries a transposon insertion in the MADS box region to generate periclinal chimeras expressing alleles with different activities. By comparing the phenotype, development and expression patterns of these chimeras we show that expression of DEF in L1 makes a major contribution to morphology in whorl 2, irrespective of the allele. By contrast L1 expression is largely unable to rescue whorl 3, possibly because of a non-autonomous inhibitor of DEF activity in this whorl.     

Low levels of allozyme variability in the threatened species Antirrhinum subbaeticum and A. pertegasii (Scrophulariaceae): implications for conservation of the species

AIMS: This study was designed to compare levels of genetic variation and its partitioning in three related species of Antirrhinum, A. subbaeticum, A. pertegasii and A. pulverulentum, and to check the hypothesis that species with small total population size have lower levels of genetic variability than those with bigger ones. This information should contribute to the development of conservation strategies of rare endemic species of Antirrhinum. METHODS: One hundred and seventy-seven plants were screened for variability at 14 allozyme loci by means of horizontal starch gel. Parameters of genetic diversity, and its partitioning, were calculated. An indirect estimate of gene flow was based on the equation: Nm = (1 - GST)/4GST. KEY RESULTS: Genetic variabilities in A. subbaeticum and A. pertegasii were found to be the lowest known for the genus, the within-population genetic diversity being correlated with population size in both species. The distribution of genetic diversity is strikingly different among species, with 85 % of the total variation distributed among populations in A. subbaeticum, 6 % in A. pertegasii and 23 % in A. pulverulentum. Estimated levels of gene flow were negligible for A. subbaeticum (0.04), high for A. pertegasii (3.92), and substantial for A. pulverulentum (0.83). Genetic and geographic distances were negatively correlated in A. pertegasii, whereas no significant correlation was found in the other two species. CONCLUSIONS: Levels of total genetic diversity agree with the hypothesis that species with small total population size have lower levels of genetic variability than those with bigger ones. Strategies for the conservation of the species are recommended, such as preservation of natural populations and avoidance of possible causes of threat, as well as ex situ preservation of seeds, reinforcement of small populations of A. subbaeticum with plants or seeds from the same population, and avoidance of translocations among populations.     

ROSINA (RSI), a novel protein with DNA-binding capacity, acts during floral organ development in Antirrhinum majus

Petal and stamen identity of the Antirrhinum majus flower is under the genetic control of the floral homeotic gene DEFICIENS (DEF). To isolate factors involved in the regulation of DEF gene activity, a promoter segment of this B-function gene, containing cis-acting regulatory elements, was used to identify the novel trans-acting factor ROSINA (RSI). RSI does not show an extended similarity with any gene product present in the database. Rather RSI constitutes a protein that contains domains similar to known proteins from organisms of different phyla. The capacity of RSI to bind a sequence element of the DEF promoter, its spatial and temporal expression pattern together with the phenotype of RSI-RNAi interference plants as well as RSI over-expression in Arabidopsis thaliana suggest that RSI is a putative regulator of DEF gene activity in A. majus.