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Philip M. Gilmartin - One of the best experts on this subject based on the ideXlab platform.
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Agrobacterium -mediated transformation systems of Primula vulgaris
Plant methods, 2018Co-Authors: Sadiye Hayta, Mark A. Smedley, Wendy Harwood, Philip M. GilmartinAbstract:Genetic transformation is a valuable tool and an important procedure in plant functional genomics contributing to gene discovery, allowing powerful insights into gene function and genetically controlled characteristics. Primulaceae species provide one of the best-known examples of heteromorphic flower development, a breeding system which has attracted considerable attention, including that of Charles Darwin. Molecular approaches, including plant transformation give the best opportunity to define and understand the role of genes involved in floral heteromorphy in the common primrose, Primula vulgaris, along with other Primula species. Two transformation systems have been developed in P. vulgaris. The first system, Agrobacterium-mediated vacuum infiltration of seedlings, enables the rapid testing of transgenes, transiently in planta. GUS expression was observed in the cotyledons, true leaves, and roots of Primula seedlings. The second system is based on Agrobacterium tumefaciens infection of pedicel explants with an average transformation efficiency of 4.6%. This transformation system, based on regeneration and selection of transformants within in vitro culture, demonstrates stable transgene integration and transmission to the next generation. The two transformation systems reported here will aid fundamental research into important traits in Primula. Although, stable integration of transgenes is the ultimate goal for such analyses, transient gene expression via Agrobacterium-mediated DNA transfer, offers a simple and fast method to analyse transgene functions. The second system describes, for the first time, stable Agrobacterium-mediated transformation of Primula vulgaris, which will be key to characterising the genes responsible for the control of floral heteromorphy.
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Genetic architecture and evolution of the S locus supergene in Primula vulgaris.
Nature plants, 2016Co-Authors: Jonathan M. Cocker, Jonathan Wright, Margaret A. Webster, Mark Mcmullan, Sarah Dyer, David Swarbreck, Mario Caccamo, Cock Van Oosterhout, Philip M. GilmartinAbstract:Darwin's studies on heterostyly in Primula described two floral morphs, pin and thrum, with reciprocal anther and stigma heights that promote insect-mediated cross-pollination. This key innovation evolved independently in several angiosperm families. Subsequent studies on heterostyly in Primula contributed to the foundation of modern genetic theory and the neo-Darwinian synthesis. The established genetic model for Primula heterostyly involves a diallelic S locus comprising several genes, with rare recombination events that result in self-fertile homostyle flowers with anthers and stigma at the same height. Here we reveal the S locus supergene as a tightly linked cluster of thrum-specific genes that are absent in pins. We show that thrums are hemizygous not heterozygous for the S locus, which suggests that homostyles do not arise by recombination between S locus haplotypes as previously proposed. Duplication of a floral homeotic gene 51.7 million years (Myr) ago, followed by its neofunctionalization, created the current S locus assemblage which led to floral heteromorphy in Primula. Our findings provide new insights into the structure, function and evolution of this archetypal supergene.
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On the origins of observations of heterostyly in Primula
The New phytologist, 2015Co-Authors: Philip M. GilmartinAbstract:In 1862, Charles Darwin published his landmark study on the different forms of flower in Primula; he coined the term distyly and subsequently expanded his studies to other species, including those with tristyly. Darwin is widely recognized as the first to study pin and thrum flowers in Primula, and to provide an explanation for the functional significance of the two floral morphs. Our laboratory is pursuing the genes that underpin floral heteromorphy in Primula, work influenced by Darwin's observations. One day, while appreciating a print of Primula vulgaris from William Curtis' Flora Londinensis, I was struck by the fact that I was looking at images of dimorphic Primula flowers captured in a late-1700s copper-plate engraving that predated Darwin's observations by over 70 yr. This realization triggered a journey into archives of botanical texts, herbals and florilegia from the 16(th) to 19(th) Centuries, and correspondence archives, in search of earlier documents that could have influenced Darwin and the origins of an idea. Darwin was not the first to observe floral heteromorphy in Primula, but he was the first to realize the significance of the two floral morphs. Darwin's insight and exposition of purpose have underpinned all consequent work on the subject.
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Molecular characterization of DNA sequences from the Primula vulgaris S-locus
Journal of experimental botany, 2005Co-Authors: Iain W. Manfield, Vassily K. Pavlov, Holly Cook, Florian Hummel, Philip M. GilmartinAbstract:Primula species provide possibly the best known examples of heteromorphic flower development and this breeding system has attracted considerable attention, including that of Charles Darwin. However, despite considerable recent advances in molecular genetics, nothing is known about the molecular basis of floral heteromorphy. The first molecular marker for the Primula S-locus is reported here. This DNA sequence was identified by random amplification of polymorphic DNA (RAPD)-PCR, further defined as a sequence characterized amplified region (SCAR) marker, and subsequently shown to correspond to a restriction fragment length polymorphism (RFLP) that is linked to the thrum allele of the Primula S-locus. The sequence of 8.8 kb of genomic DNA encompassing this thrum-specific RFLP is presented. Analysis of this DNA reveals a highly repetitive sequence structure similar to that found at the S-locus in other species; it also contains sequences similar to elements of a Gypsy-like retrotransposon. The identification of a specific DNA sequence associated with the thrum allele of the Primula S-locus provides the first molecular probe with which to investigate the molecular basis of heteromorphic flower development in Primula.
B Raimondi - One of the best experts on this subject based on the ideXlab platform.
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seed germination and conservation of endangered species from the italian alps physoplexis comosa and Primula glaucescens
Biological Conservation, 2004Co-Authors: Bruno Enrico Leone Cerabolini, Rossella De Andreis, Roberta M Ceriani, Simon Pierce, B RaimondiAbstract:Abstract Physoplexis comosa and Primula glaucescens , endemic to calcareous grasslands of the Lombardy Prealps, northern Italy, are protected under Annex IV of European Community Directive 92/43/EEC. Seed germination dynamics of these species were studied with the aim of producing appropriate germination protocols for use in ex situ conservation. Seeds were collected from Monte Barro (922 m a.s.l.) and for Primula glaucescens from three additional sites in a more central part of its range (1217–2521 m a.s.l.). Physoplexis comosa germinated only in sterile in vitro conditions, in which germination percentages were extremely low without a supply of gibberellic acid (GA3; 1.7%); the optimum GA3 concentration was 100 mg l −1 (> 90% germination), with lower germination at higher doses of 250–500 mg l −1 . Primula glaucescens germinated in both sterile and non-sterile conditions and, although total germination was not affected by GA3 concentration, doses of 10–500 mg l −1 acted equally to prompt earlier and more rapid germination. Primula glaucescens from Monte Barro was characterised by slower and poorer germination than from central sites.
C. R. Lovell - One of the best experts on this subject based on the ideXlab platform.
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FS11.1 Primula obconica – a falling allergen
Contact Dermatitis, 2008Co-Authors: Maureen Connolly, J Mc Cune, E. Dauncey, C. R. LovellAbstract:Objective: We believe the incidence of Primula contact allergic dermatitis has fallen since the introduction of primin-free Primula onto the European market and thus our study aims were twofold. Firstly to see if the incidence of Primula contact allergic dermatitis was truly on the decline and secondly to confirm the presence and document retailers’ knowledge and awareness of primin-free Primula in the UK. Methods: A questionnaire was sent to 22 contact dermatitis departments throughout the UK and Ireland looking at the number of primin positive patch tests in the years 1995/96, 1998, 2000 and 2002 compared with the total number of patch tests. 10 seed suppliers and 12 plant retailers were asked to complete a telephone survey. Results: We showed a significant fall in the yearly incidence of contact allergy to primin from 0.785% in 1995/96 to 0.457% in 2002. This downward trend was statistically significant (p = 0.001). The telephone survey showed 90% of seed suppliers were aware that the older varieties of P. obconica could cause an allergic reaction whereas only 60% of them were aware that new primin-free varieties were now being bred. 50% of suppliers were in fact selling these primin-free varieties with 60% of them stocking a primin-free variety exclusively. 90% of retailers were not currently selling any variety of P. obconica. Conclusion: Our study shows that the incidence of Primula contact dermatitis is falling. The overall trend is moving towards primin-free varieties provided they continue to be horticulturally viable long term.
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Contact dermatitis due to hardy Primula species and their cultivars.
Contact dermatitis, 2001Co-Authors: Carolyn G. Aplin, C. R. LovellAbstract:For more than 100 years, it has been widely held that of all the Primula species to cause allergic contact dermatitis, P. obconica is the most likely culprit, particularly in Northern Europe. The main sensitizer is primin, but other sensitizers are reported. Other species are rarely implicated, but we suspect from our discussions with Primula growers that this is probably a consequence of under-reporting. Moreover, cutaneous reactions to other hardy Primula species and cultivars would appear to be milder and may sometimes be associated with a state of tolerance after repeated handling. With the co-operation of members of the Midland and West Section of the National Auricula and Primula Society of the UK, 462 questionnaires were sent out to members and 316 replies were returned (response rate 68.4%) with data on 320 growers. 84 out of a total of 320 (26.25%) attributed a cutaneous reaction to Primula species, whereas 236 (73.75%) reported no reaction. Of the 84 who reported reactions, 48 suspected P. auricula, 34 P. obconica, 10 P. vulgaris, 5 P. allionii, 3 P. marginata and 2 P. forrestii 19 believed that they had reacted to 2 or more species of Primula. Our study suggests that Primula species other than P. obconica may elicit dermatitis more frequently than previously recorded.
Gelsomina Fico - One of the best experts on this subject based on the ideXlab platform.
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Phytochemistry of European Primula species.
Phytochemistry, 2017Co-Authors: P. Colombo, Guido Flamini, Graziella Rodondi, Claudia Giuliani, Laura Santagostini, Gelsomina FicoAbstract:Abstract The genus Primula is the largest among the Primulaceae and is widespread mainly in the cold and temperate regions of the Northern Hemisphere. Since the beginning of the Twentieth century, several studies on the phytochemical composition of different species of Primula have been carried out. The main constituents examined were tissue and epicuticular flavonoids and saponins, which are of therapeutic significance. Only in recent years studies of the volatiles emitted by leaves and flowers have been carried out as well, but they are restricted to a small number of species. Only a few authors have documented the morphology and function of glandular trichomes in relation to the production of flavonoids and volatile organic compounds (VOCs). The use of Primula in folk medicine is described in the literature. Investigation of the biological and pharmacological activities of Primula are reported. This study aims at providing a collection of publications on the genus Primula along with a critical revision of literature data. It focuses on the possible taxonomic significance of the secondary metabolites and on their ecological role as attractors for pollinators and deterrents against herbivores and parasites, in order to build the base for further studies.
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Primula latifolia Lapeyr. and Primula vulgaris Hudson flavonoids
Natural product research, 2014Co-Authors: P. Colombo, Guido Flamini, Gelsomina FicoAbstract:Three flavonoids were isolated from the leaf MeOH extracts of Primula latifolia Lapeyr. and Primula vulgaris Hudson collected from Italian Alps: rutin (1) and kaempferol 3-neohesperidoside (2) from P. latifolia, and kaempferol 3-β-O-glucopyranosyl-(1 → 2) gentiobioside (3) from P. vulgaris. The structures were assigned on the basis of their 1H and 13C NMR data, including those derived from 2D NMR, as well as on HPLC–MS results. This article is the first to report on P. vulgaris tissue flavonoids after Harborne's study in 1968 and the first work ever on these compounds from P. latifolia.
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Comparative phytochemical and morphological analyses of three Italian Primula species
Phytochemistry, 2007Co-Authors: Gelsomina Fico, Guido Flamini, Graziella Rodondi, Daniele Passarella, Franca TomèAbstract:Abstract The taxonomy of alpine Primula species has long been in dispute because of high morphologic variability and several hybridisations. In Primula species, the trichome height and the colour of hair-tips are usually indicated as diacritic characters, but in our experience this is not adequate. The present study, focused on Primula auricula, Primula daonensis and Primula hirsuta, therefore proposes the use of other morphologic trichome parameters (size and dimensional ratio of stalk, neck and gland head). Phytochemical investigations about the flavonoid composition (epicuticular and vacuolar) of leaves, as taxonomic markers, have also been performed. We report the isolation and identification of two new flavonol glycosides, isorhamnetin 3-O-(2,6-di-O-β- d -glucopyranosyl-β- d- glucopyranoside) (1) and kaempferol 3-O-(2-O-α- l -rhamnopyranosyl-6-O-β- d -xylopyranosyl-β- d -glucopyranoside) (2) and of eight known flavonoids. Size and dimensional ratio of the three trichome elements (stalk, neck and glandular head) are typical for each species analysed. The flavonoid profile well characterise the entities under study. Three different profiles have been obtained with both vacuolar and epicuticular flavonoids. The morphologic and phytochemical markers proposed in this work seem to be parameters which significatively discriminate the species under study.
Hai-fei Yan - One of the best experts on this subject based on the ideXlab platform.
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the complete chloroplast genome of a rare candelabra primrose Primula stenodonta Primulaceae
Conservation Genetics Resources, 2017Co-Authors: Cai-yun Zhang, Hai-fei Yan, Tongjian Liu, Gang HaoAbstract:The whole chloroplast genome (cp genome) of Primula stenodonta was sequenced using an Illumina platform. The cp genome is 150,785 bp in size, and exhibits a typical bicyclic and quadripartite structure, including a pair of inverted repeat regions (IRs) of 25,182 bp that are separated by large and small single copy regions of 82,682 and 17,739 bp, respectively. A total of 131 genes were identified, including 86 protein-coding genes (PCG), 37 tRNA genes, and eight ribosomal RNA (rRNA) genes. Among them, 18 genes occur in IRs, containing seven PCG, seven tRNA genes and four rRNA genes. The overall AT content of P. stenodonta cp genome is 62.9%, and the corresponding values of the LSC, SSC and IR regions are 65, 69.8 and 57%, respectively. The phylogenetic analysis suggested that the sampled Primula species formed a monophyletic clade, and P. stenodonta is closely related to Primula poissonii.
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DNA Barcoding Evaluation and Its Taxonomic Implications in the Species-Rich Genus Primula L. in China
PloS one, 2015Co-Authors: Hai-fei Yan, Yun-jiao Liu, Xiu-feng Xie, Cai-yun Zhang, Gang HaoAbstract:The genus Primula is extremely diverse in the east Himalaya-Hengduan Mountains (HHM) in China as a result of rapid radiation. In order to overcome the difficulty of morphological classification of this genus, we surveyed three plastid regions (rbcL, matK, and trnH-psbA) and two nuclear markers (ITS and ITS2) from 227 accessions representing 66 Primula species across 18 sections, to assess their discriminatory power as barcodes. We found that ITS alone or combined with plastid regions showed the best discrimination across different infrageneric ranks and at species level. We suggest rbcL + matK + ITS as the first choice at present to barcode Primula plants. Although the present barcoding combination performed poorly in many closely related species of Primula, it still provided many new insights into current Primula taxonomy, such as the underlying presence of cryptic species, and several potential improper taxonomic treatments. DNA barcoding is one useful technique in the integrative taxonomy of the genus Primula, but it still requires further efforts to improve its effectiveness in some taxonomically challenging groups.
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Comparative transcriptome analysis and marker development of two closely related Primrose species (Primula poissonii and Primula wilsonii).
BMC genomics, 2013Co-Authors: Lu Zhang, Hai-fei YanAbstract:Primula species are important early spring garden plants with a centre of diversity and speciation in the East Himalaya-Hengduan Mountains in Western China. Studies on population genetics, speciation and phylogeny of Primula have been impeded by a lack of genomic resources. In the present study, we sequenced the transcriptomes of two closely related primrose species, Primula poissonii and Primula wilsonii, using short reads on the Illumina Genome Analyzer platform. We obtained 55,284 and 55,011 contigs with N50 values of 938 and 1,085 for P. poissonii and P. wilsonii, respectively, and 6,654 pairs of putative orthologs were identified between the two species. Estimations of non-synonymous/synonymous substitution rate ratios for these orthologs indicated that 877 of the pairs may be under positive selection (Ka/Ks > 0.5), and functional enrichment analysis revealed that significant proportions of the orthologs were in the categories DNA repair, stress resistance, which may provide some hints as to how the two closely related Primula species adapted differentially to extreme environments, such as habitats characterized by aridity, high altitude and high levels of ionizing radiation. It was possible for the first time to estimate the divergence time between the radiated species pair, P. poissonii and P. wilsonii; this was found to be approximately 0.90 ± 0.57 Mya, which falls between the Donau and Gunz glaciation in the Middle Pleistocene. Primers based on 54 pairs of orthologous SSR-containing sequences between the two Primula species were designed and verified. About half of these pairs successfully amplified for both species. Of the 959 single copy nuclear genes shared by four model plants (known as APVO genes), 111 single copy nuclear genes were verified as being present in both Primula species and exon-anchored and intron-spanned primers were designed for use. We characterized the transcriptomes for the two Primula species, and produced an unprecedented amount of genomic resources for these important garden plants. Evolutionary analysis of these two Primula species not only revealed a more precise divergence time, but also provided some novel insights into how differential adaptations occurred in extreme habitats. Furthermore, we developed two sets of genetic markers, single copy nuclear genes and nuclear microsatellites (EST-SSR). Both these sets of markers will facilitate studies on the genetic improvement, population genetics and phylogenetics of this rapidly adapting taxon.