Patterns of genetic and phenotypic variation in Iris haynei and I. atrofusca (Iris sect. Oncocyclus = the royal irises) along an ecogeographical gradient in Israel and the West Bank

Iris haynei and I. atrofusca are two closely related narrow endemics distributed vicariously along an ecogeographical north-south gradient in Israel and the West Bank. To obtain baseline information of the taxonomic status, conservation and population history of these taxa, we investigated patterns of phenotypic variation and the partitioning of genetic variation within and among populations using dominant random amplified polymorphic DNA (RAPD) markers. Multivariate (principal components analysis) and taxonomic distance analyses based on morphometric traits from eight populations revealed no unambiguous separation into two distinct groups. Results of genetic analyses for nine populations differed only slightly when either allele- or marker-based approaches were employed. Mean within-population diversity was high (0.258 for Nei's expected heterozygosity), but there was no significant relationship between genetic diversity and either population size or latitude. Although the range-wide estimate of GST ( approximately 0.20) revealed relatively high differentiation among populations this value was inflated because of a small, but significant, component of molecular variance among regions viz. taxa ( approximately 5%). Limited long-distance dispersal capabilities in conjunction with a linearized habitat distribution are proposed to contribute to the approximate isolation by distance pattern observed. It also appears that extant populations are currently deviating from equilibrium conditions because of primary divergence of a formerly more widespread ancestral population. Given the absence of deep genetic and phenotypic subdivision among northern (I. haynei) vs. central/southern (I. atrofusca) populations, we argue for a revision of their species status. Nonetheless, we recommend conservation attention to these geographically differentiated segments as separate management units, which can be seen as an instructive example of incipient species formation.     

Morphological variation of the Oncocyclus irises (Iris: Iridaceae) in the southern Levant

Morphological traits of Iris section Oncocyclus (Siems.) Baker in the southern Levant (Israel, Jordan, The Palestinian Authority and Sinai/Egypt) were analysed in order to clarify taxonomic relationships among taxa and the validity of diagnostic characters. Floral and vegetative characters were measured in 42 populations belonging to nine species during the peak of the flowering season in 1998–2000. Pearson's Coefficient of Racial Likelihood (CRL) was used to calculate morphological distances between populations. Twelve of the measured populations, distributed along the north-south aridity gradient in Israel, were further explored for morphological changes along the gradient. Cluster analysis revealed two major clusters: the first includes most of the dark-coloured Iris populations, with populations of I. petrana Dinsmore and I. mariae W. Barbey forming a subcluster; the second consists of all the light-coloured populations but also some dark-coloured populations. Pearson's CRL and geographical distance were significantly correlated among the dark-coloured populations. Along the geographical gradient, flower, stem and leaf size traits decrease towards the south, probably as an adaptation to aridity. This suggests that natural selection promoted the differences between populations. Almost no discrete phenotypic groups exist within the Oncocyclus species of the southern Levant except for variation in the floral colours. Most of the suggested diagnostic characters proved unreliable in that they varied continuously across populations. The taxonomical difficulties encountered in this study reflect the special evolutionary state of the Oncocyclus irises as a group in the course of speciation.  © 2002 The Linnean Society of London. Botanical Journal of the Linnean Society , 2002, 139 , 369–382.     

Concordance and discordance: a tale of two hybrid zones in the pacific coast irises (Iridaceae)

Chloroplast DNA (cpDNA) markers were developed that provided markers unique to a species or that delimited a large area within a species. These markers were then followed across two hybrid zones: Iris douglasiana/Iris∗∗∗ innominata, and Iris chrysophylla/Iris tenax. In each case the cline in haplotype frequency was compared to the cline for a morphologically based hybrid index. In all three transects across the I. douglasianall. innominata hybrid zone, the cpDNA cline was displaced 1-2 km relative to the morphologically defined hybrid zone; the displacement was not found in the other hybrid zone. The observed displacement represents introgression of cpDNA from ∗∗∗I. douglasiana into ∗∗∗I. innominata. It may be that the I. douglasiana/I.∗∗∗ innominata hybrid zone has shifted in recent time, leaving the slowly dispersing chloroplast DNA behind. The populations known as Iris thompsonii do not form a phylogenetic species and are best viewed as products of hybridization between ∗∗∗I. douglasiana and ∗∗∗I. innominata.     

Isolation and characterization of 10 microsatellite loci from Iris hexagona (Iridaceae)

Ten microsatellite loci isolated from Iris hexagona are described. All these loci are polymorphic, with three to 13 alleles across 24 individuals from a single natural population. Heterozygosity ranged from 0.125 to 0.870. Three loci depart significantly from Hardy–Weinberg equilibrium in our test population. The test population shows significant heterozygote deficiency in these and two other loci. Three loci exhibit significant linkage disequilibrium. These loci will be utilized to investigate patterns of genetic variation in the species throughout the Florida peninsula.     

Molecular and morphological evidence reveals hybridisation between two endemic cyprinid fishes

The geographic occurrence of moggel Labeo umbratus × mud fish Labeo capensis hybrids and the threat of hybridisation to the genetic integrity of L. umbratus were investigated. Genetic evidence from mtDNA cytb and nDNA s7 intron sequence data indicated probable interspecific hybridisation in two impoundments, Hardap Dam (Orange River Basin, Namibia) and Darlington Dam (Sundays River Basin, Eastern Cape, South Africa). Some putative hybrids were morphologically identifiable on account of their meristic and morphometric intermediacy to the parent species. Human activities appear to drive the observed hybridisation, either through dam construction and direct stocking of L. umbratus or via translocation of Labeo capensis and the previously isolated Orange River lineage of L. umbratus into the southern-flowing systems by inter-basin water transfers. It is recommended that further translocation of fish from these areas should be avoided.     

玉蝉花繁殖生态学研究

通过野外调查、人工授粉和室内种子萌发实验等,对天目山玉蝉花（Iris ensata）种群繁殖生态特征进行了研究。结果显示：玉蝉花自然种群呈聚集型分布,种群更新主要依赖贡献率为0.756的无性繁殖。人工同瓣和异瓣自花授粉座果率分别为10%和20%,结实数分别为0和4±1（n=2）粒/果;同株和异株异花授粉座果率分别为80%和100%,结实数分别为59±7（n=8）和64±9（n=10）粒/果,自交主要来自同株异花授粉,而同株花异熟、雌雄异位和异向等可有效降低自交。自然套袋不结实,不存在无融合生殖。花粉/胚珠比1277±270（n=10）,辅证玉蝉花为兼性异花授粉、异花授粉植物。有效授粉昆虫为蜜蜂（Apismellifica）,访花频率为0.019次.花-1.h-1,单只蜜蜂回访同朵花的比率为3.5%。自然种群立地种子数估算值为368粒/m2,种子宿存,具非深度生理休眠,萌发需光,低温层积60d萌发率达94.9±0.7%。玉蝉花种群繁殖适应对策为k对策,具备自我维持机制,但现正受人为干扰。     

Use of RAPD analysis in devising conservation strategies for the rare and endangered Grevillea scapigera (Proteaceae)

Extensive human impact in south western Australia has resulted in a high incidence of rarity throughout the highly endemic flora of the region. Grevillea scapigera (Proteaceae) is a typical example, with 27 plants (represented by four extant populations) remaining in the wild. In order to devise an appropriate strategy for the conservation of this species, its population genetics were studied using RAPD analysis, which enabled the discrimination of individual plants and the detection of a relatively high amount of variability (V = 0.32) within G. scapigera. This variability was found to be evenly distributed within the plants analysed despite the clear distinction between most populations (87% of the variability being attributable to single plant difference and 13% to population difference). Finally, RAPD analysis was used to select a small group of plants that captured maximum genetic variability to be used in the recovery program of the species. Because of the low genetic difference between populations, the mixing of these selected plants during the recovery process should not create genetic imbalances. The methods used in this study provide a useful model for future projects involving the recovery of rare flora.     

Population genetic variation in rare and endangered Iliamna (Malvaceae) in Virginia

Random amplified polymorphic DNA (RAPD) markers were used as input for an analysis of molecular variance (AMOVA), homogeneity of molecular variance analysis (HOMOVA), and cluster analysis to describe the population genetic structure of Iliamna corei, a federally endangered plant located only in Virginia, and I. remota , a rare plant in Virginia, Indiana, and Illinois. The analysis was performed to help clarify the taxonomic relationship between the two closely related species. We analysed four clones in the only known population of I. corei , breeding stock derived from seeds originating from the population site, and three I. remota populations in Virginia. Eighty-five percent of screened primers revealed DNA polymorphisms in Iliamna. Ninety-nine informative markers were generated using seven primers. No significant statistical differences (at P = 0.05) in RAPD variation was found between species (24% of variance) using the AMOVA procedure. However, within species/among populations (31 % of the variance) and within populations (45% of the variance) there were significant differences (P < 0.002). An unweighted paired group method using arithmetic averages (UPGMA) cluster analysis showed the federally endangered I. corei population to be genetically distinct from the apparently recently introduced (in Virginia: ∼ 100 ybp) I. remota. The lack of significant differences from the AMOVA and the high number shared bands between I. corei and I. remota suggest that I. corei may be more appropriately classified as a subspecies of I. remota. Iliamna corei plants in the natural population were genetically similar to one another while the I. corei breeding stock plants and I. remota plants were genetically relatively diverse.     

Molecular and morphometric variation in chromosomally differentiated populations of the grasshopper Sinipta dalmani (Orthopthera: Acrididae)

Sinipta dalmani is an Argentine grasshopper whose chromosome polymorphisms have been widely studied through cytogenetic, morphometric, and fitness component analyses. The present work analysed molecular and morphometric variation in seven chromosomally differentiated populations from Entre Rios and Buenos Aires provinces to analyse population structure. Molecular studies were performed studying RAPD loci and morphometric analyses were carried out measuring five morphometric traits. Genetic variability was high in all studied populations and was characterized by a decrease in H as a function of latitude and temperature. Both conventional F(ST) analysis and Bayesian approach for dominant marker showed that there were significant genetic differences among all populations, between provinces, and among populations within provinces. Entre Rios populations showed higher mean numbers of migrants per generation as well as low genetic differentiation and high gene flow with almost all populations whereas Buenos Aires populations may be considered as a result of a more recently colonization. There is considerable morphometric variation between populations and this variation correlates with latitude and temperature. Our results suggest that selection contributes to phenotypic differentiation among populations by moulding the differences in trait means whereas genetic drift is responsible for differences in the matrix of variance-covariance. The gene flow detected is insufficient to prevent phenotypic and chromosome divergences.     

Isozyme diversity in Iris Cristata and the threatened glacial endemic I. Lacustris (Iridaceae)

Iris cristata and I. lacustris differ markedly in geographic distribution, glacial history of current ranges, and ecology. We hypothesized that I. cristata, a widespread species of unglaciated regions of eastern North America, would exhibit genetic diversity typical of other widespread plant species, whereas the threatened I. lacustris, which occupies glaciated habitats on Great Lakes shorelines, would display little genetic variation. Iris lacustris lacked detectable polymorphisms in 18 isozyme loci, although we found evidence of possible incomplete gene silencing in four additional loci in some populations. In contrast, I. cristata was polymorphic at 73% of 15 loci examined, with an average of three alleles per locus. Genetic diversity (H_e) was 0.231. All species-level and population-level estimates of genetic diversity were higher than averages for plants having comparable life history traits. Nearly 98% of the total genetic diversity in I. cristata was apportioned within populations, and heterozygosity and fixation estimates suggest a high level of outcrossing in this species (t = 1.265). The long-lived perennial habit and high outcrossing rate in stable populations are proposed as factors contributing to high genetic diversity in I. cristata. The data are consistent with an hypothesis of a recent origin of I. lacustris from a very limited I. cristata gene pool exacerbated by repeated bottlenecks and founder effects as I. lacustris populations were displaced by lake-level changes over the past 11 000 yr.     

Pollination of Oncocyclus irises (Iris: Iridaceae) by night-sheltering male bees

Irises in the section Oncocyclus (Siems.) Baker ( IRIS: Iridaceae) grow throughout the Middle East and have large and dark-coloured flowers but no nectar reward available to flower visitors. Consequently, no reward-collecting pollinators have been observed visiting the flowers during daytime. The only visitors are solitary male bees ( Eucera spp.: Apidae) that enter the flowers at dusk and stay there overnight. Here we describe the mating system of Oncocyclus irises, and the role of night-sheltering male bees in their pollination system. Pollen viability in I. haynei on Mt. Gilboa was very high (>90%) throughout all floral life stages. Stigmas were receptive in buds and in open flowers, but not in older ones. Self-pollination yielded no fruits in three species, confirming complete self-incompatibility in Oncocyclus irises. On average, 1.9 flowers were visited by each male bee before it settled for the night in the last one. Moreover, Iris pollen was present on the dorsal side of 38.8% of males caught sheltering in flower models mounted near an I. atrofusca population, indicating that pollen is transferred between flowers by night-sheltering solitary male bees. We have surveyed 13 flowering populations of six Oncocyclus species for the presence of night-sheltering male bees as well as for fruit set. We found a positive correlation, indicating that sexual reproduction in Oncocyclus irises is dependent on night-sheltering solitary male bees. Based on their complete self-incompatibility, the absence of nectar-collecting visitors during the day, and the transfer of pollen grains by the night-sheltering solitary male bees, we conclude that fertilization of Oncocyclus irises is totally dependent on pollination by night-sheltering solitary male bees.     

Author and type of the name Iris cengialti (Iridaceae)

Abstract

A. Kerner is correctly the author of the name Iris cengialti previously ascribed to Ambrosi, and consequently the type of the species is here selected, as lectotype, among Kerner's original material kept in WU.     

Isolation and characterization of polymorphic microsatellites in Iris ensata (Iridaceae)

• Premise of the study: Microsatellite primers were developed in Iris ensata (Iridaceae) to provide polymorphic markers for further studies into population genetics. • Methods and Results: Thirteen polymorphic microsatellite loci were isolated from I. ensata. These loci were successfully amplified in two natural populations of I. ensata from eastern China (Longwangshan, Zhejiang Province) and northeastern China (Jinchuan, Jilin Province). There was no significant linkage disequilibrium found for any pair of loci. These loci contained between two and 12 alleles per locus across all 48 individuals of I. ensata. The number of alleles per locus varied from two to 10 at the population level and the observed and expected heterozygosities ranged from 0.167 to 0.958 and from 0.284 to 0.853, respectively. • Conclusions: These loci showed high levels of polymorphism and could be used to study the population genetic structure, genetic relationships, and phylogeography of I. ensata.     

Utility of DNA barcoding to identify rare endemic vascular plant species in Trinidad

The islands of the Caribbean are considered to be a “biodiversity hotspot.” Collectively, a high level of endemism for several plant groups has been reported for this region. Biodiversity conservation should, in part, be informed by taxonomy, population status, and distribution of flora. One taxonomic impediment to species inventory and management is correct identification as conventional morphology‐based assessment is subject to several caveats. DNA barcoding can be a useful tool to quickly and accurately identify species and has the potential to prompt the discovery of new species. In this study, the ability of DNA barcoding to confirm the identities of 14 endangered endemic vascular plant species in Trinidad was assessed using three DNA barcodes (matK, rbcL, and rpoC1). Herbarium identifications were previously made for all species under study. matK, rbcL, and rpoC1 markers were successful in amplifying target regions for seven of the 14 species. rpoC1 sequences required extensive editing and were unusable. rbcL primers resulted in cleanest reads, however, matK appeared to be superior to rbcL based on a number of parameters assessed including level of DNA polymorphism in the sequences, genetic distance, reference library coverage based on BLASTN statistics, direct sequence comparisons within “best match” and “best close match” criteria, and finally, degree of clustering with moderate to strong bootstrap support (>60%) in neighbor‐joining tree‐based comparisons. The performance of both markers seemed to be species‐specific based on the parameters examined. Overall, the Trinidad sequences were accurately identified to the genus level for all endemic plant species successfully amplified and sequenced using both matK and rbcL markers. DNA barcoding can contribute to taxonomic and biodiversity research and will complement efforts to select taxa for various molecular ecology and population genetics studies.     

Maternal effects on early juvenile traits inIris pumila (Iridaceae)

The variation in the maternal effects on a number of the earlyIris pumila seedling traits expressed under high and low light levels in a growthroom was studied. A reciprocal crossing design was used for the establishment of 20 full-sib families. Each family consisted of two maternal groups differing only in genotype used as a seed parent. Maternal variation was estimated from the variance between maternal groups within sibships. Two parameters for maternal variation were computed: the maternal effects coefficient m² and the coefficient of maternal variation CV_M. Maternal effects significantly accounted for the phenotypic variation in seed mass and most of the early seedling traits. Light conditions prevailing during the seedling development markedly influenced overall structure of maternal correlations. Implications of maternal correlations on the integration of juvenile phenotype have also been discussed.     

Genetic diversity of the endangered endemic Agave victoriae-reginae (Agavaceae) in the Chihuahuan Desert

Long-lived perennials are a species-rich, ecologically important component of the North American deserts, yet we know little about their genetic structure, information important for their conservation. Agave victoriae-reginae is an endemic of the Chihuahuan Desert of northern Mexico that is endangered by collection for the ornamental trade. We examined levels and patterns of variation at ten polymorphic allozyme loci in ten populations representing the range of the species. Levels of genetic variation (mean H_e= 0.335) and differentiation (mean F_ST = 0.236) were high. Phenetic clustering suggested the existence of at least three distinct groups of populations. If this pattern of variation is representative of other long-lived desert perennials, it may explain the species richness of this group and will pose a real challenge to gene conservation efforts.     

Molecular phylogenetics of the juno irises,Iris subgenus Scorpiris (Iridaceae), based on six plastid markers

Relationships among the roughly 55 species of Iris subgenus Scorpiris have been studied. A matrix of six plastid DNA regions (matK, rpl14‐rps8 spacer, infA‐rpl36 spacer, trnE‐trnT spacer, trnL intron and trnL‐F spacer) was produced from 57 accessions (52 taxa) and analysed with both parsimony and Bayesian methods. Five major clades are identified, of which four have strong geographical correlations, whereas the fifth corresponds to Iris section Physocaulon. In our results, several species are placed with species not previously considered to be related, although, in some cases, there are morphological characters that suggest that these newly indicated relationships are reasonable. For some of the other oddly grouped species, we can only assume that remarkable parallelisms in morphology have occurred or hybridization is involved. Presently, with plastid DNA as our only comprehensive data resource, we are not able to evaluate more thoroughly these more puzzling associations of species. © 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167 , 281–300.     

Floral development of Iris decora Wall. (Iridaceae)

The outer tepal and stamen primordia arise as secondary primordia on the outer tepal-stamenprimordia, which are formed on the floral apex. The inner tepal primordia are formed directly on the floral apex. All the floral appendages are initiated in the second tunica layer and are homologous with regard to their origin and early development. A short perianth tube is formed as a result of intercalary growth in the common bases of the tepal primordia. The intercalary growth in the fused bases of the floral appendages elevates the peripheral zone. The floral apex thus appears as a shallow cup. Further intercalary growth results in the formation of an inferior ovary. The ovules are initiated as outgrowths on placental ridges from the lateral ovary wall, the trilocular appearance being the result of secondary cohesion of the parietal placentae.     

Pollinator shifts as triggers of speciation in painted petal irises (Lapeirousia: Iridaceae)

Background and Aims

Adaptation to different pollinators has been hypothesized as one of the main factors promoting the formation of new species in the Cape region of South Africa. Other researchers favour alternative causes such as shifts in edaphic preferences. Using a phylogenetic framework and taking into consideration the biogeographical scenario explaining the distribution of the group as well as the distribution of pollinators, this study compares pollination strategies with substrate adaptations to develop hypotheses of the primary factors leading to speciation in Lapeirousia (Iridaceae), a genus of corm-bearing geophytes well represented in the Cape and presenting an important diversity of pollination syndromes and edaphic preferences.

Methods

Phylogenetic relationships are reconstructed within Lapeirousia using nuclear and plastid DNA sequence data. State-of-the-art methods in biogeography, divergence time estimation, character optimization and diversification rate assessments are used to examine the evolution of pollination syndromes and substrate shifts in the history of the group. Based on the phylogenetic results, ecological factors are compared for nine sister species pairs in Lapeirousia.

Key Results

Seventeen pollinator shifts and ten changes in substrate types were inferred during the evolution of the genus Lapeirousia. Of the nine species pairs examined, all show divergence in pollination syndromes, while only four pairs present different substrate types.

Conclusions

The available evidence points to a predominant influence of pollinator shifts over substrate types on the speciation process within Lapeirousia, contrary to previous studies that favoured a more important role for edaphic factors in these processes. This work also highlights the importance of biogeographical patterns in the study of pollination syndromes.     

ISSR variation in the endemic and endangered plant Cycas guizhouensis (Cycadaceae)

BACKGROUND AND AIMS: Cycas guizhouensis (Cycadaceae) is a rare and endangered species endemic to the southwest of China. An investigation was undertaken into the genetic variation of wild populations. METHODS: ISSR markers were used to determine the genetic variation within and between 12 extant populations of this species. KEY RESULTS: Low genetic diversity (at population level, P = 14.21 %, H(E) = 0.0597; at species level, P = 35.90 %, H(T) = 0.1082) and a high degree of differentiation among populations (G(ST) = 0.4321) were detected. CONCLUSIONS: This genetic structure is considered to be due to the combined effects of slow biochemical evolution, genetic drift, inbreeding and limited gene flow between populations. Based on these findings, strategies are proposed for the genetic conservation and management of the species.