Identification of the Key Regulatory Genes Involved in Elaborate Petal Development and Specialized Character Formation in Nigella damascena (Ranunculaceae).

Petals can be simple or elaborate, depending on whether they have lobes, teeth, fringes, or appendages along their margins, or possess spurs, scales, or other types of modifications on their adaxial/abaxial side, or both. Elaborate petals have been recorded in 23 orders of angiosperms and are generally believed to have played key roles in the adaptive evolution of corresponding lineages. The mechanisms underlying the formation of elaborate petals, however, are largely unclear. Here, by performing extensive transcriptomic and functional studies on Nigella damascena (Ranunculaceae), we explore the mechanisms underlying elaborate petal development and specialized character formation. In addition to the identification of genes and programs that are specifically/preferentially expressed in petals, we found genes and programs that are required for elaborate rather than simple petal development. By correlating the changes in gene expression with those in petal development, we identified 30 genes that are responsible for the marginal/ventral elaboration of petals and the initiation of several highly specialized morphological characters (e.g., pseudonectaries, long hairs, and short trichomes). Expression and functional analyses further confirmed that a class I homeodomain-leucine zipper family transcription factor gene, Nigella damascena LATE MERISTEM IDENTITY1 (NidaLMI1), plays important roles in the development of short trichomes and bifurcation of the lower lip. Our results not only provide the first portrait of elaborate petal development but also pave the way to understanding the mechanisms underlying lateral organ diversification in plants.

[Ecology suitability study of Trollius chinensis].

Based on the distribution information of 110 samples and 55 environmental factors, Maxent model was used to predict the ecology suitability regions of Trollius chinensis. The study aims at providing theory basis for the cultivation of T. chinensis. The results showed that the Maxent model prediction result was good (AUC>0.9) and the main factors effecting the ecology suitability regions of T. chinensis were precipitation in July, standard deviation of seasonal variation of temperature, annual mean temperature, precipitation in August and altitude. The ecology suitable regions of T. chinensis mainly concentrated in Shanxi, Hebei, east of Inner Mongolia, west of Jilin and Liaoning, north of Shaanxi, south of Ningxia, east and south of Gansu, and east of Qinghai. The results indicated that except for traditional distribution regions, north of Shaanxi, south of Ningxia, east and south of Gansu, and east of Qinghai could selected as the regions for cultivation of T. chinensis. It provides theory basis for selecting suitable regions to cultivate T. chinensis.

Composición y distribución de hepáticas (Marchantiophyta) en un intervalo altitudinal en la Cordillera Oriental de Colombia / Composition and distribution of liverworts (Marchantiophyta) in a continuous altitudinal range on the Cordillera Oriental of Colombia

Resumen La composición de especies de hepáticas sobre los Andes del Norte varía con la altitud en relación con diferentes factores bióticos (vegetación) y abióticos (temperatura, humedad y luminosidad). Para probar esta afirmación se determinó la diversidad y distribución de las Marchantiophytas en un rango altitudinal entre los 2 400 y 3 400 m sobre la Cordillera Oriental colombiana, estableciendo cómo éstas varían entre altitudes y zonas de vida con respecto a los factores abióticos. Se realizaron muestreos cada 200 m y se registraron datos ambientales durante un año. Se encontraron 162 especies, la composición de hepáticas varió a lo largo del gradiente altitudinal con un gran número de especies únicas en cada altitud. Se encontró la máxima diversidad en los 3 000 m, con una dominancia del hábito folioso y de especies epifitas, mientras que la mayor riqueza de familias y géneros se presentó entre los 3 200 y 3 400 m. Anoplolejeunea conferta presentó la mayor cobertura en la zona así como el mayor valor del índice de valor de importancia (IVI) y distribución altitudinal más amplia, ubicándose desde los 2 400 hasta 3 000 m en cuatro diferentes sustratos. Las hepáticas se distribuyeron altitudinalmente de manera diferencial, con una diversidad beta alta (0.864) debido al recambio altitudinal de especies, con más del 30 % de disimilaría en la composición de especies cada 200 m altitudinales, dependiendo principalmente de cambios en la luminosidad. Consecuentemente, la temperatura, humedad y luminosidad y su relación con la vegetación son determinantes en la diversidad y distribución de las hepáticas en la Cordillera Oriental de los Andes colombianos.

Abstract The species composition of liverworts varies with altitude in the Northern part of the Andes due to the relationship of biotic (vegetation) and abiotic factors (temperature, humidity, and sunlight brightness). In order to test this affirmation we determined the diversity, species composition, abundance, and distribution of species of Marchantiophyta in an altitudinal range from 2 400 to 3 400 m on the Colombian Cordillera Oriental, and established how these vary with altitude and life-zones, regarding abiotic factors. Samples, taken every 200 m in the altitudinal range, and environmental data were registered during a year. We found 162 species, the composition of liverworts varied throughout the altitudinal gradient with a high number of unique species in each altitude. The maximum diversity was found at 3 000 m, along with a dominance of leafy habit and epiphytic species, while the higher richness of families and genera was found between 3 200 and 3 400 m. Anoplolejeunea conferta had the largest value of coverage in the zone, the highest value of importance value index (IVI), and the widest altitudinal distribution, from 2 400 to 3 000 m in four different substrates. The liverworts were distributed differently with altitude, we found a high beta diversity (0.864) due to the replacement of species, with more that 30 % of dissimilarity in species composition every 200 altitudinal meters, mainly depending on sunlight brightness variation. Accordingly, we found that abiotic factors like temperature, humidity, and sunlight brightness and its relationship vegetation are determinant in the diversity and altitudinal distribution of liverworts in the study area. Rev. Biol. Trop. 66(2): 559-570. Epub 2018 June 01.

Cytogeography of the subalpine marsh marigold polyploid complex (Caltha leptosepala s.l., Ranunculaceae).

PREMISE OF THE STUDY: Unrecognized variation in ploidy level can lead to an underestimation of species richness and a misleading delineation of geographic range. Caltha leptosepala (Ranunculaceae) comprises a complex of hexaploids (6x), rare nonaploids (9x), and dodecaploids (12x), all with unknown distributions. We delineate the geographic distribution and contact zones of the cytotypes, investigate morphologies of cytotypes and subspecies, and discuss the biogeography and evolutionary history of the polyploid complex. METHODS: Using cytologically determined specimens as reference, propidium iodide flow cytometry was performed on silica-dried samples and herbarium specimens from across the range of C. leptosepala s.l. Genome size estimates from flow cytometry were used to infer cytotypes. A key morphological character, leaf length-to-width ratio, was measured to evaluate whether these dimensions are informative for taxon and/or cytotype delimitation. KEY RESULTS: Dodecaploids were more northerly in distribution than hexaploids, and a single midlatitude population in the Northern Rockies yielded nonaploids. Genome size estimates were significantly different between all cytotypes and between hexaploid subspecies. Leaf length-to-width ratios were significantly different between subspecies and some cytotypes. CONCLUSIONS: Caltha leptosepala presents clear patterns of cytotype distribution at the large scale. Marked differences in morphology, range, and genome size were detected between the hexaploid subspecies, C. leptosepala subsp. howellii in the Cascade-Sierra axis and C. leptosepala subsp. leptosepala in the Rockies. Sympatry between cytotypes in the Cascades and a parapatric distribution in the Northern Rockies suggest unique origins and separate lineages in the respective contact zones.

Genetics of flower development in Ranunculales - a new, basal eudicot model order for studying flower evolution.

Contents 361 I. 361 II. 362 III. 363 IV. 364 V. 364 Acknowledgements 365 References 365 SUMMARY: Ranunculales, the sister group to all other eudicots, encompasses species with a remarkable floral diversity, which are currently emerging as new model organisms to address questions relating to the genetic architecture of flower morphology and its evolution. These questions concern either traits only found in members of the Ranunculales or traits that have convergently evolved in other large clades of flowering plants. We present recent results obtained on floral organ identity and number, symmetry evolution and spur formation in Ranunculales species. We discuss benefits and future prospects of evo-devo studies in Ranunculales, which can provide the opportunity to decipher the genetic architecture of novel floral traits and also to appraise the degree of conservation of genetic mechanisms involved in homoplasious traits.

The rise of angiosperm-dominated herbaceous floras: Insights from Ranunculaceae.

The rise of angiosperms has been regarded as a trigger for the Cretaceous revolution of terrestrial ecosystems. However, the timeframe of the rise angiosperm-dominated herbaceous floras (ADHFs) is lacking. Here, we used the buttercup family (Ranunculaceae) as a proxy to provide insights into the rise of ADHFs. An integration of phylogenetic, molecular dating, ancestral state inferring, and diversification analytical methods was used to infer the early evolutionary history of Ranunculaceae. We found that Ranunculaceae became differentiated in forests between about 108-90 Ma. Diversification rates markedly elevated during the Campanian, mainly resulted from the rapid divergence of the non-forest lineages, but did not change across the Cretaceous-Paleogene boundary. Our data for Ranunculaceae indicate that forest-dwelling ADHFs may have appeared almost simultaneously with angiosperm-dominated forests during the mid-Cretaceous, whereas non-forest ADHFs arose later, by the end of the Cretaceous terrestrial revolution. Furthermore, ADHFs were relatively unaffected by the Cretaceous-Paleogene mass extinction.

Plant Functional Diversity Can Be Independent of Species Diversity: Observations Based on the Impact of 4-Yrs of Nitrogen and Phosphorus Additions in an Alpine Meadow.

Past studies have widely documented the decrease in species diversity in response to addition of nutrients, however functional diversity is often independent from species diversity. In this study, we conducted a field experiment to examine the effect of nitrogen and phosphorus fertilization ((NH4)2 HPO4) at 0, 15, 30 and 60 g m-2 yr-1 (F0, F15, F30 and F60) after 4 years of continuous fertilization on functional diversity and species diversity, and its relationship with productivity in an alpine meadow community on the Tibetan Plateau. To this purpose, three community-weighted mean trait values (specific leaf area, SLA; mature plant height, MPH; and seed size, SS) for 30 common species in each fertilization level were determined; three components of functional diversity (functional richness, FRic; functional evenness, FEve; and Rao's index of quadratic entropy, FRao) were quantified. Our results showed that: (i) species diversity sharply decreased, but functional diversity remained stable with fertilization; (ii) community-weighted mean traits (SLA and MPH) had a significant increase along the fertilization level; (iii) aboveground biomass was not correlated with functional diversity, but it was significantly correlated with species diversity and MPH. Our results suggest that decreases in species diversity due to fertilization do not result in corresponding changes in functional diversity. Functional identity of species may be more important than functional diversity in influencing aboveground productivity in this alpine meadow community, and our results also support the mass ratio hypothesis; that is, the traits of the dominant species influenced the community biomass production.

Contrasting relationships between species diversity and genetic diversity in natural and disturbed forest tree communities.

• This study aimed to reveal species-genetic diversity correlations (SGDCs) and their underlying mechanisms in natural and disturbed forests. • A community survey and molecular analyses were carried out to compare species diversity (SD), the genetic diversity of the dominant tree species Euptelea pleiospermum (GD), the altitudinal patterns of SD and GD, SGDC, genetic differentiation (F(ST) ), community divergence (F(ST) -C), effective population size (N(e) ), and recent migration rate between mountain riparian forests along the Yandu (natural) and Nan (disturbed) rivers. • In natural forests, both SD and GD showed a unimodal altitudinal pattern and GD was positively correlated with SD, whereas a unimodal pattern and positive SGDC were not found in the disturbed forests. SD and F(ST) at the natural sites were higher than those at the disturbed sites. However, there were no significant differences in GD, F(ST) -C, N(e) or recent migration rate between the natural and disturbed sites. • A correlation between the patterns of SD and GD along a geographical gradient (e.g. altitude) is an important driver of positive SGDC. The absence of positive SGDC in the disturbed forests may result from reduced SD but unaffected GD, indicating nonparallel changes in SD and GD. This study furthermore cautions against generalizations about changes in SD and GD following disturbance.

The shape of things to come: woodland herb niche contraction begins during recruitment in mesic forest microhabitat.

Natural abundance is shaped by the abiotic requirements and biotic interactions that shape a species' niche, yet these influences are rarely decoupled. Moreover, most plant mortality occurs during early life stages, making seed recruitment critical in structuring plant populations. We find that natural abundance of two woodland herbs, Hexastylis arifolia and Hepatica nobilis, peaks at intermediate resource levels, a pattern probably formed by concurrent abiotic and biotic interactions. To determine how this abundance patterning reflects intrinsic physiological optima and extrinsic biotic interactions, we translocate adults and seeds to novel locations across experimentally extended abiotic gradients. These experiments indicate that the plant distributions probably reflect biotic interactions as much as physiological requirements, and that adult abundance provides a poor indication of the underlying niche requirements. The positive response exhibited by adult transplants in the wettest conditions is offset by increased fungal attack on buried seeds consistent with peak natural abundance where soil moisture is intermediate. This contraction of niche space is best described by Connell's model--species are limited by physiological tolerances where resources are low and biotic interactions where resources are high.

An experimental test of well-described vegetation patterns across slope aspects using woodland herb transplants and manipulated abiotic drivers.

*The ubiquitous transition of plant communities across slope aspects is a well-described, but rarely tested, ecological dynamic. Aspect position is often used as a proxy for microclimate changes in moisture, light and temperature, but these abiotic drivers are seldom decoupled and very rarely manipulated across slope aspects. *To investigate the mechanisms and demographic stages driving the observed distribution patterns of two woodland herbs in the southeastern USA, seeds and adults were transplanted across north- and south-facing slopes, and moisture and light were experimentally manipulated. *Stage- and species-specific abiotic responses resulted in similar landscape-level patterning for Hexastylis arifolia and Hepatica nobilis, but the underlying abiotic drivers were unique. Adult rather than seed survival best explained the natural distributions across slope aspects, and Hexastylis arifolia was limited by higher temperature, whereas Hepatica nobilis was limited by lower soil moisture. *The stage- and species-specific responses indicated that the use of slope aspect to explain plant distributions not only obfuscates explanatory mechanisms, but probably undermines the portability of results. As abiotic drivers, not topographical proxies, are projected to shift with global climate change, distribution research requires direct abiotic data in association with key demographic stages rather than topographical proxies.

[Nutrient contents in Kingdonia uniflora ramet and their relations to heterogeneous environment on Taibai Mountains].

Taking the guerrilla type clonal plant Kingdonia uniflora at different altitudes in Taibai Mountain National Forest Park of Shaanxi Province, China as test material, the C, N, P, K and Mg contents in different vegetative organs of its ramet were measured. The results showed that ramet age, altitude, and their interaction had no significant effects on the total C content in ramet leaf, rhizome, and root (P > 0.05), but different effects on the total N, total P, K and Mg contents in these vegetative organs, suggesting that ramet could regulate the allocation of nutritive elements in its vegetative organs through physiological metabolism. At high altitude (3000-3100 m), ramet age had significant effects on the K content in all vegetative organs and the N content in rhizome and root (P < 0.05). All the test nutrient contents were significantly correlated with each other at all altitudes, except at middle altitude (2800-2900 m) where the ramet K content had no significant correlations with its N, P, and Mg contents. It was suggested that physiological plasticity contributed more to the performances of clonal plants at high altitude than at low altitude. Powerful physiological plasticity could help K. uniflora to effectively utilize heterogeneous resources, realize population expansion, and open up new habitat at high altitude. It was likely that the physiological plasticity of K. uniflora responding to heterogeneous habitats was formed in its long-term evolvement.

Ovule morphogenesis in Ranunculaceae and its systematic significance.

BACKGROUND AND AIMS: Ranunculaceae has a prominent phylogenetic position in Ranunculales which appears at the base of eudicots. The aims of the present paper are to reveal the features of ovule morphogenesis in different taxa and gain a better understanding of the systematics of Ranunculaceae. METHODS: Flowers of 17 species from three subfamilies, nine tribes and 16 genera of Ranunculaceae, at successive developmental stages, were collected in the wild and studied with a scanning electron microscope. KEY RESULTS: The integuments in the unitegmic ovules in Helleborus, Ranunculus and Oxygraphis, as well as the inner integuments in the bitegmic genera, initiate annularly and eventually become cup-shaped. However, the integuments in the unitegmic ovules in Anemone and Clematis, as well as the outer integuments in the bitegmic genera, arise semi-annularly and eventually become hood-shaped. Different kinds of appendages appear on the ovules during development. In Coptis of subfamily Coptidoideae, a wrap-shaped appendage arises outside the ovule and envelopes the ovule entirely. In the genera of subfamily Thalictroideae and tribe Anemoneae of subfamily Ranunculoideae, appendages appear on the placenta, the funicle or both. In tribe Helleboreae of subfamily Ranunculoideae, an alary appendage is initiated where the integument and the funicle join and becomes hood-shaped. CONCLUSIONS: Ovule morphogenesis characteristics are significant in classification at the levels of subfamilies and tribes. The initiation patterns of the integuments and the development of appendages show diversity in Ranunculaceae. The present observations suggest that the bitegmic, hood-shaped outer integument and endostomic micropyle are primitive while the unitegmic, cupular-shaped outer integument and bistomic micropyle are derivative.

Differential responses of the freshwater wetland species Juncus effusus L. and Caltha palustris L. to iron supply in sulfidic environments.

Sulfur pollution can lead to serious problems in freshwater wetlands, including phosphorus eutrophication and sulfide toxicity. We tested the effects of anaerobic iron-rich groundwater discharge in fens, simulated by iron injection, on two characteristic species (Juncus effusus and Caltha palustris) in a sulfidic environment. Biomass production of C. palustris roots showed an optimum response to the combined addition of iron and sulfide, with highest values at intermediate concentrations of both substances. Iron deficiency apparently occurred at low iron concentrations, while at high iron concentrations, growth was decreased. For J. effusus, in contrast, no toxic effects were found of both iron and sulfide. This could be explained by larger radial oxygen loss (ROL) of J. effusus and could not be explained by differences in phosphorous concentrations. The results of our experiments confirm that iron-rich groundwater discharge has the potential to affect vegetation composition through toxicity modification in sulfidic environments.

[Identification study of different Flos Trollii by IR].

OBJECTIVE: To establish a method to identify different Flos Trollii with IR spectra and to provide a new technique for their science estimation and quality identification. METHODS: Their IR spectra were obtained by the method of intuitionistic analysis-selecting point to normalization. RESULTS: Differences on the shape, the position, the height and the relative intensity of absorption peaks were shown in IR spectra of the ten samples. CONCLUSIONS: Different Flos Trollii can be identified using IR spectra obtained by the method of intuitionistic analysis-selecting point to normalization. This new method can be used to study the identification features of the influence factors of Traditional Chinese Medicine quality.

[Analysis on age structure and dynamics of Kindonia uniflora populations].

Kindonia uniflora is a perennial clone herbaceous plant, and also, a native endangered plant in China. This paper studied its age structure, life table and survivorship curve in different habitats in Taibai mountain area. The results indicated that the age structure and dynamics of K. uniflora populations in the Betula utilis forest at altitude 2500-2700 m, in the Abies fargesii forest at altitude 2700-2900 m, and in the Larix chinensis forest at altitude 2900-3100 m had the similar pattern and developing tendency. The number of younger ramets at 1-2 years old or older than 5 years was less, and the number of ramets at 3-5 years old was the highest in the age structures. The negative values of dx (dead number), qx (mortality rate) and Kx (Killing rate) in the life table showed the increasing rate of the population sizes during the age stage. The survivorship curve of K. uniflora populations in different habitats belonged to Deevey C after 3-5 years old. The mortality rate of populations during 5-10 years stage was higher, and was stable after 10 years old. As for the characters of asexual propagation and clone growth, the rhizomes of the populations were in humus of soil, and developed and expanded as guerilla line style. During growth season, only one leaf grew above ground at every inter-node, and the population growth and development were rarely influenced by external factors. The forest communities, such as Betula utilis, Abies fargesii and Larix chinensis forest, in which K. uniflora populations lived, were at middle or higher mountain, where there were rarely disturbance from human being. Therefore, the habitats for K. uniflora populations to live were relatively stable. As the altitude increased, the disturbances from human being became less, the density of K. uniflora populations increased, the life cycle expanded, the peak of population death delayed, and the population living strategy changed to adapt to the habitats. K. uniflora populations preferred to live in cool climate, thick humus of soil and under shaded forest, therefore, the habitat of communities of Abies fargesii, Larix chinensis forest at altitude 2700-3100 m were better than that in Betula utilis forest. The growth and development of K. uniflora populations depended on the habitats; therefore, the communities in which K. uniflora population lived should be conserved firstly. The in-situ conservation of the populations should be encouraged.

[Individual and modular biomass dynamics of Kingdonia uninflora population in Qinling Mountain].

The individual and modular biomass dynamics of Kingdonia uninflora populations in Qinling Mountain were studied systematically. They could be expressed by Logistic equation. The ratio of leaf to bud biomass of population decreased with its increasing age, whereas the ratio of the root system to the subterranean stem biomass increased. The individual and each modular biomass of different populations at different altitudes were significant different (P < 0.05), which were larger at altitude 2700-2900 m than at the lower and higher altitudes. The rate of leaf biomass decreased as altitude rose, whereas that of the subterranean stem increased. In the favorable area, the rate of root system biomass was less, whereas the bud biomass was bigger compared with other habitats. The individual growth and modular biomass accumulation were closely related to habitat condition.

Developing fruit direct post-floral morphogenesis in Helleborus niger L.

In fertilized flowers of Helleborus niger L., the sepals (the showy elements of the perianth at anthesis) grow, spread, and turn green, and the peduncles elongate. These processes did not proceed to completion when the pistils were removed at the bud stage, but could be restored by the application of plant growth regulators. Cytokinins and gibberellins stimulated the formation of well-developed chloroplasts in, and spreading of, the sepals; the gibberellin, GA3, and the auxin, 4-chloroindole-3-acetic acid, promoted peduncle elongation. In fruit-bearing flowers, on the other hand, paclobutrazol, an inhibitor of gibberellin biosynthesis, reduced chlorophyll formation in the sepals, reversed sepal spreading, and inhibited peduncle elongation. Of the endogenous growth regulators in developing fruit, the following cytokinins were identified: zeatin, dihydrozeatin, N6-(2-isopentenyl)adenine and their ribosides and 9-glucosides. Zeatin riboside drastically increased in abundance (about 200 times), shortly after fertilization, when chlorophyll accumulation in the sepals occurred at the fastest rate, and remained the most prominent identified cytokinin until seed ripening.

Genes from the APETALA3 and PISTILLATA lineages are expressed in developing vascular bundles of the tuberous rhizome, flowering stem and flower Primordia of Eranthis hyemalis.

In Arabidopsis thaliana expression of the B-class MADS-box genes APETALA3 (AP3) and PISTILLATA (PI) is confined to petals and stamens but in other plant species these genes are also transcribed in non-flower tissues; in Solanum tuberosum they are transcribed specifically in vascular bundles leading to petals and stamens. Transcription analysis of B-class genes in Eranthis hyemalis using reverse transcribed in situ PCR revealed that both AP3 and PI are expressed in developing vascular bundles in the tuberous rhizome, flowering stem and floral primordia. In addition, AP3 and PI transcripts are also found in stems and leaves. These results suggest a more complex role of B-class genes in Eranthis and possible involvement in the development of vascular tissue.

Development of STS and CAPS markers for identification of three tall larkspurs (Delphinium spp.).

One cleaved amplified polymorphic sequence (CAPS) and nine sequence tagged site (STS) markers were developed for identifying tall larkspur (Delphinium spp.) plants in three species based on the DNA sequence of known species-specific RAPD markers. Four STS markers were used for identification of Delphinium occidentale, three STS markers for Delphinium barbeyi, and one CAPS and two STS markers for Delphinium glaucum. One hundred sixty-six individual plants collected at 19 locations in the western U.S.A. were tested using the STS and CAPS markers. Over 95% of the D. occidentale plants contained all four D. occidentale specific STS markers, whereas the remaining plants contained three of the four STS markers. Approximately 97% of D. barbeyi plants contained all three D. barbeyi specific STS markers, and the rest had two of the three STS markers. A small percentage of D. barbeyi plants contained one D. occidentale specific STS marker. Hybrid populations were characterized as having more D. occidentale specific than D. barbeyi specific STS markers, suggesting that the three hybrid populations are composed not of F1 hybrid plants of the parental species but of segregating offspring of different generations from original hybrids. This set of STS and CAPS markers for larkspur species should be useful in classification of unknown plant materials and the identification of hybrid populations.