Low genetic diversity and functional constraint of miRNA genes participating pollen–pistil interaction in rice

Key message

In this study, we sequenced and analyzed the expression and evolution of rice miRNA genesparticipating pollen-pistil interaction that is crucial to rice yield.

Abstract

Pollen–pistil interaction is an essential reproductive process for all flowering plants. While microRNAs (miRNAs) are important noncoding small RNAs that regulate mRNA levels in eukaryotic cells, there is little knowledge about which miRNAs involved in the early stages of pollen–pistil interaction in rice and how they evolve under this conserved process. In this study, we sequenced the small RNAs in rice from unpollinated pistil (R0), pistil from 5 min and 15 min after pollination, respectively, to identify known and novel miRNAs that are involved in this process. By comparing the corresponding mRNA-seq dataset, we identified a group of miRNAs with strong negative expression pattern with their target genes. Further investigation of all miRNA loci (MIRNAs) across 1083 public rice accessions revealed significantly reduced genetic diversity in MIRNAs with strong negative expression of their targets when comparing to those with little or no impact on targets during pollen–pistil interaction. Annotation of targets suggested that those MIRNAs with strong impact on targets were pronounced in cell wall related processes such as xylan metabolism. Additionally, plant conserved miRNAs, such as those with functions in gibberellic acid, auxin and nitrate signaling, were also with strong negative expression of their targets. Overall, our analyses identified key miRNAs participating pollen–pistil interaction and their evolutionary patterns in rice, which can facilitate the understanding of molecular mechanisms associated with seed setting.

     

Discordant patterns of morphological and genetic divergence in the closely related species Schizanthus hookeri and S.?grahamii (Solanaceae)

In this study I have examined the patterns of morphological and genetic differentiation between two species of the Andean genus Schizanthus that differ in their pollination and mating systems. Schizanthus hookeri has a bee pollination syndrome and is strongly dependent on pollinators for seed set. In contrast, S.?grahamii has a hummingbird pollination syndrome and exhibits late autonomous selfing. Southern populations of the latter species have red flowers (reddish morph), while northern populations have yellow (yellowish morph) or pink flowers (pinkish morph). I used two noncoding chloroplast DNA (cpDNA) regions to investigate the genetic affinities between S.?hookeri and the three morphs of S.?grahamii. I also performed intra- and interspecific crosses to assess whether gene flow between species was possible. Phylogenetic analyses supported the existence of two differentiated clades that did not match currently accepted taxonomic classification. Accordingly, genetic distance did not correlate significantly with morphological distance. No fruits were produced from interspecific crosses, and there were no individuals with intermediate morphology that could indicate current and frequent hybridization events between species.?I propose that the discordance between cpDNA data and conventional taxonomy could be explained by parallel evolution, or alternatively, by a very sporadic hybridization.     

Cellular localization and levels of pectins and arabinogalactan proteins in olive (Olea europaea L.) pistil tissues during development: implications for pollen–pistil interaction

Cell wall components in the pistil are involved in cell–cell recognition, nutrition and regulation of pollen tube growth. The aim of this work was to study the level, whole-organ distribution, and subcellular localization of pectins and arabinogalactan proteins (AGPs) in the olive developing pistil. Western blot analyses and immunolocalization with fluorescence and electron microscopy were carried out using a battery of antibodies recognizing different types of pectin epitopes (JIM7, JIM5, LM5, and LM6) and one anti-AGPs antibody (JIM13). In the olive pistil, highest levels of acid esterified and de-esterified pectins were observed at pollination. Moreover, pollination was accompanied by a slight decrease of the galactose-rich pectins pool, whereas arabinose-rich pectins were more abundant at that time. An increased expression of AGPs was also observed during pollination, in comparison to the pistil at the pre-anthesis stage. After pollination, the levels of pectins and AGPs declined significantly. Inmunofluorescence localization of pectins showed their different localization in the olive pistil. Pectins with galactose residues were located mainly in the cortical zones of the pistil, similar to the neutral pectins, which were found in the parenchyma and epidermis. In turn, the neutral pectins, which contain arabinose residues and AGPs, were localized predominantly in the stigmatic exudate, in the cell wall of secretory cells of the stigma, as well as in the transmitting tissue of the pistil during the pollination period. The differences in localization of pectins and AGPs are discussed in relation to their roles during olive pistil developmental course.     

Length conversions and mass–length relationships of five forage-fish species in the California current ecosystem

Length-measurement conversions and seasonal mass–length relationships (MLR) for Pacific herring Clupea pallasii, northern anchovy Engraulis mordax, Pacific sardine Sardinops sagax, Pacific mackerel Scomber japonicus and jack mackerel Trachurus symmetricus in the California Current are presented. The conversions between total (L_T), fork (L_F,) and standard lengths (L_S) should facilitate comparisons of data across disciplines and institutions. These equations resulted from an analysis of measurements spanning 14 years and the western seaboard of North America, from the north end of Vancouver Island to the USA–Mexico border. Major-axis regressions were used to calculate reciprocal length-measurement conversions (e.g., L_T to L_S and L_S to L_T) and generalised linear models and ordinary least-squares models were used to create MLRs that account for seasonal variations. The MLR models indicated seasonal differences for all species except C. pallasii, for which there was no multi-season data. Discrepancies between these and published models were examined, along with the suitability and benefit of the various types of models used for length-measurement conversion and MLRs.     

Cryptohelops menaticus – a new genus and species of the tribe Helopini (Coleoptera: Tenebrionidae) from the Palaeocene of Menat (France)

A new genus and species, Cryptohelops menaticus gen. et sp. n., are described from the Palaeocene of Menat (France). The new genus belongs to the subtribe Helopina of the tribe Helopini (subfamily Tenebrioninae of Tenebrionidae) based on the coarse and dense punctation of the hypomera, shape of the prosternum and metepisterna, a similar appearance to the native genera Helops and Stenohelops and the structure of the epipleura. The new genus also resembles Raiboscelis and Entomogonus by having the protibiae excised along the base of the inner side. The new genus is the oldest representative of the tribe Helopini. Four species of Helops sensu lato previously described from the Oligocene (H. wetteravicus K. Heyden et L. Heyden, 1865) and Miocene (H. atticus Redtenbacher in Ungern, 1867; H. meissneri Heer, 1847 and H. molassicus Heer, 1883) lack important diagnostic characters and should be regarded as members of the tribe Helopini with uncertain generic attribution.     

Length–weight relationships for five species of Chimaerids and two species of Rhinochimaerids to the west of Ireland

Length–weight regressions for five chimaerid species, Chimaera monstrosa Linnaeus, 1758, Chimaera opalescens Luchetti, Iglesias & Sellos, 2011, Hydrolagus affinis de Brito Capello, 1868, Hydrolagus pallidus Hardy & Stehmann, 1990 and Hydrolagus mirabilis Collett, 1904, and two Rhinochimaerid species, Rhinochimaera atlantica Holt & Byrne, 1909 and Harriotta raleighana Goode & Beane, 1895, are calculated from data collected during a series of deepwater surveys conducted by the Irish Marine Institute from 2006 to 2009. This work was carried out on the continental slope to the west and northwest of Ireland and the northern slope of the Porcupine bank. Samples were measured fresh at the end of each haul. Due to the fragility of the tails of many of these species, length measurements for Chimaerids were made to the pre supra caudal fin, while for Rhinochimaerids they were made to the second dorsal fin. Lengths were measured to the nearest centimetre below and weights were recorded to the nearest gram. Outliers were removed from the dataset according to difference in fits (DFFITS) (Belsley et al., Regression diagnostics: Identifying influential data and sources of collinearity, John Wiley & Sons; 1980). b values for the Chimaeridae ranged from 2.915 to 3.075, while r² values ranged from .981 to .995. b values for H. raleighana and R. atlantica were 2.764 and 3.505 and r² values were .977 and .976 respectively.     

Edge effect and species–area relationships in the gall-forming insect fauna of natural forest patches in the Brazilian Pantanal

The aim of this study was to (i) measure differences in species richness between edge habitats versus interior habitats, or more precisely the edge effect, and (ii) test the species–area relationship for gall-forming insects in natural forest patches in a Brazilian floodplain (Pantanal of Mato Grosso do Sul). These patches are regionally known as capões, basically composed of woody vegetation. Twenty-seven patches were surveyed. In each patch two transects were conducted for gall sampling. One transect encircled the patches while the other was conducted in the interior of the patch, totaling 54h of sampling. Host plant and galling insect species composition differed quite characteristically between the edge and the interior of patches, but galling insect richness did not. When insect gall richness was expressed as the ratio between insect gall and host plant richness (gall per plant ratio), a weak species–area relationship was found. Our results suggest that the number of galling insects per individual plant is not affected by the size of the patch. Despite these results, the natural forest patches found in this region seem well suited for long-term studies addressing species–area relationships. With regard to herbivorous insects, these studies should be combined with research on host plant dynamics during flooding and dry seasons.     

Comparisons among populations and individuals to evaluate pollen–pistil interaction as a mechanism of reproductive interference in Taraxacum

Reproductive interference (RI), an interspecific mating interaction that reduces the fitness of at least one of the species involved, can lead to exclusive distributions in closely related species. A hypothesis previously proposed is that RI in plants may occur by ovule usurpation, in which pistils lack interspecific incompatibility and mistakenly accept heterospecific pollen, thereby losing an opportunity for conspecific pollen fertilization. However, few comparative studies have evaluated the consistency of the inferred mechanism within and among individuals and populations. We conducted hand-pollination experiments in six populations of three native Taraxacum species that suffered from different levels of RI from an alien congener, T. officinale, and compared pollen–pistil interactions among populations. We also investigated the interactions for eight individual T. japonicum plants whose response to heterospecific pollen deposition had been previously measured. Our results revealed that pollen tubes often penetrated native ovaries following heterospecific pollination in populations suffering from strong RI, whereas they seldom did in populations suffering from marginal RI. However, the relative frequency of the pollen tube penetration was not significantly related to the strength of alien RI. Not all pistils on an individual plant showed the same pollen receptivity following heterospecific pollination; rather, some accepted and some refused the pollen tubes. The relationship between pollen tube penetration following heterospecific pollination and the strength of the alien RI was also not significant among individuals. Our present results generally support the ovule usurpation hypothesis, but suggest that other factors, such as competition for pollinator services, variation in the effects of heterospecific pollen donors, and condition of the native inflorescences, might also affect the observed RI strength.

     

Pollen–pistil barriers to crossing in maize and teosinte result from incongruity rather than active rejection

Many popcorn strains cannot be fertilized by pollen of dent and flint strains although the reciprocal crosses are successful. Similarly, plants of some annual teosinte populations can fertilize maize but do not accept its pollen. Single genes or gene complexes govern these two unilateral barriers to crossing. Failure of fertilization could reflect active rejection by the pistil of pollen containing a contrasting allele (incompatibility). Alternatively, the pistil could require presence of a matching allele in pollen (congruity). To distinguish between these possibilities genetically, the receptivity to pollen having both alleles was determined. If there is active rejection, heteroallelic pollen would not be accepted; if presence of a matching allele is required, heteroallelic pollen would be accepted. In both the popcorn and teosinte crossing barrier systems, heteroallelic pollen functioned, consistent with the congruity model.     

(–)-Germacrene D receptor neurones in three species of heliothine moths: structure-activity relationships

Specificity of olfactory receptor neurones plays an important role in food and host preferences of a species, and may have become conserved or changed in the evolution of polyphagy and oligophagy. We have identified a major type of plant odour receptor neurones responding to the sesquiterpene germacrene D in three species of heliothine moths, the polyphagous Heliothis virescens and Helicoverpa armigera and the oligophagous Helicoverpa assulta. The neurones respond with high sensitivity and selectivity to (–)-germacrene D, as demonstrated by screening via gas chromatography with numerous mixtures of plant volatiles. Germacrene D was present in both host and non-host plants, but only in half of the tested species. The specificity of the neurones was similar in the three species, as shown by the "secondary" responses to a few other sesquiterpenes. The effect of (–)-germacrene D was about ten times stronger than that of the (+)-enantiomer, which again was about ten times stronger than that of (–)--ylangene. Weaker effects were obtained for (+)--ylangene, (+)--copaene, -copaene and two unidentified sesquiterpenes. The structure-activity relationship shows that the important properties of (–)-germacrene D in activating the neurones are the ten-membered ring system and the three double bonds acting as electron-rich centres, in addition to the direction of the isopropyl-group responsible for the different effects of the germacrene D enantiomers.Abbreviations -copaene (tricyclo[4.4.0.0.]dec-3-ene, 1,3-dimethyl-8-(1-methylethyl)-) - -copaene (tricyclo[4.4.0.0.]decane, 1-methyl-3-methylene-8-(1-methylethyl)-) - GC gas chromatograph - GC-MS linked gas chromatography mass spectrometry - GC-SCR linked gas chromatography single cell recording - GD germacrene D (1,6-cyclodecadiene, 1-methyl-5-methylene-8-(1-methylethyl-, E,E)) - -ylangene (tricyclo[4.4.0.0]dec-3-ene, 1,3-dimethyl-8-(1-methylethyl)-) - -ylangene (tricyclo[4.4.0.0.]decane, 1-methyl-3-methylene-8-(1-methylethyl)-)     

Pollen Morphology of the Family Zingiberaceae in China—Pollen Types and Their Significance in the Taxonomy

Pollen morphology of 89 species and 3 varieties belonging to 18 genera (out of 150 spcies in 20 genera) of Zingiberaceae in China was studied under both light microscope and scanning electron microscope. Pollen grains of Zingiberaceae are spherical, subspherical, ovoid and prolate, 36-225 μm in size, nonaperturate or aperturate (spiraperturate, porate). Pollen grains are almost not resistant to acetolysis. The wall is composed of a very thin exine and a thick intine. The exine is psilate, spinate, cerebelloid-areolate, striate, verrucate and foveolate. According to the presence or the absence of aperture and differential ornamentations, two types and six subtypes are recognized: I. The type Nonaperturate: (85 species and 3 varieties in 18 genera). Four subtypes can be recognized within the type based on the characteristics of the exine sculpture. These are: (1) The subtype Psilate, in which, the exine is nearly smooth (including: Hedychium, Curcuma, Kaempferia, Caulokaempferia coenobilis, Boesenbergia rotunda, Stahlianthus, Amomum compactum, Etingera, Hornstedtis, Rhynchanthus). (2) The subtype Spinate, which comprises two groups: (A) The group Short-spinate, pollen grains with smaller spines (Globba), (B) The group Long-spinate, pollen grains with longer spines (Alpinia, Amomum, Plagiostachys, Roscoea, Cautleya, Boesenbergia fallax, Caulokaempferia yunnanensis). (3) The subtype Cereblloid-areolate, pollen grains of which are spherical or subspherical, with cerebelloid sculpture (Zingiber Sect. Zingiber). (4) The subtype Striate, pollen grains of which are prolate or oliveshaped, and striate (Zingiber Setc. Cryptanthium). II. The type Aperturate, in which pollen grains are acetilysis-resistant and possess distinct apertures (mixed colpate-porate or forate), including two subtypes: (1)The subtype Mixed colpate and Porate. Pollen grains are both 3-colpate and 1-3-porate, and usually with one long spiral, two short (straight or slightly curved) colpi and 1-3-poris. The exine is verrucate or not, nearly sinuolate (Costus speciosus, C. tonkinensis, C. lacerus). (2) The subtype porate, whose grains are 6-8-porate and exine is foveolate (Costus megalobractea). The taxonomic significance of the pollen types in the family Zingiberaceae is also discussed.     

Dynamism and context‐dependency in diversification of the megadiverse plant genus Solanum (Solanaceae)

Explosive radiations—substantial increases in net species diversification—have been considered one of the most intriguing diversification patterns across the Tree of Life, but the subsequent change, movement, and extinction of the constituent lineages make radiations hard to discern or understand as geological time passes. We used the megadiverse angiosperm genus Solanum L. (Solanaceae), with ca. 1200 currently accepted species distributed worldwide in a wide array of habitats, to explore these patterns on a global scale. We synthesized phylogenetic and distributional data for this ongoing radiation to show how dispersal events and past climatic changes have interacted to shape diversification. We find that, despite the vast diversity of Solanum lineages in the Neotropics, lineages in the Old World are diversifying more rapidly. This recent increase in diversification coincides with a long‐distance dispersal event from the Neotropics to regions where major climatic changes were taking place. Two separate groups of Solanum have migrated and established in Australia, but only the arid‐adapted lineages underwent significant increases in diversification rate, as they were able to adapt to the continent's long‐term climatic trend towards seasonally dry and arid biomes (a pattern observed in the diversification of other arid‐adapted groups). Our findings provide a clear example of how successful colonization of new areas and niches can—but does not always—drive explosive diversifications.     

Intergeneric pollen – megagametophyte relationships of conifers in vitro

 Germinating pollen from larch (Larix occidentalis), Sitka spruce (Picea sitchensis) and white pine (Pinus monticola) were co-cultured with megagametophytes dissected from cones of other genera (Pseudotsuga menziesii, Larix×eurolepis and Pinus monticola). Pollen was presented to megagametophytes possessing archegonia which were either alive, degenerating or dead. In addition, pollen was presented to fertilized megagametophytes and to megagametophytes that had been cut in half. Megagametophyte penetration by pollen tubes and male gamete release into archegonia were verified by serial sections of glycomethacrylate-embedded specimens. Pollen tubes penetrated through any part of the apex of the megagametophyte. Division of the body cell into the two gametes was regularly observed. Delivery of gametes was confirmed between spruce and larch. Pollen tubes also penetrated fertilized megagametophytes, dead or degenerating archegonia as well as wounded and/or cut surfaces. This demonstrates the inability of the male gametophyte to optimize its mating efforts, since it is unable to differentiate between healthy and unhealthy archegonia. The megagametophyte cells are unable to optimize male selection. They may produce secretions of a generally attractive nature, as pollen is attracted to the apex of the megagametophyte, but archegonia themselves do not produce pollen-specific signals of either a promotive or inhibitory nature. These results open new avenues for the development of novel breeding strategies where natural breeding barriers may be bypassed. Received: 19 March 1998 / Accepted: 29 April 1998     

Characterization of mode of reproduction in Commiphora wightii [(Arnot) Bhandari] reveals novel pollen–pistil interaction and occurrence of obligate sexual female plants

Guggul [Commiphora wightii (Arnot) Bhandari], a polygamous woody tree valued for its medicinal oleoresin gum rich in guggulsterone, is reported to reproduce via sporophytic apomixis. Details about its natural diversity, and mode and extent of sexual reproduction are, however, scanty. Therefore, a comprehensive investigation of guggul reproduction was made employing histology, controlled pollination, flow cytometry and RAPD analyses of progeny to assess the occurrence and extent of sexual reproduction. We report the discovery of obligate sexual female plants of guggul through these studies. Also, we document a unique pollen–pistil incompatibility that prevents all but one pollen tube growth into the style to effect fertilization. Consequently, obligate sexual female plants produced single-seeded fruit although each flower contains four ovules. In apomictic plants bearing more than one seed per fruit, at most only one seed was of sexual origin. Further, flow cytometric analysis conclusively demonstrated that endosperm development occurs either autonomously or following triple fusion. Autonomous endosperm development was invariably associated with endoreduplication, a unique feature of apomixis in guggul. Despite predominance of apomixis, a low frequency of sexual reproduction was found to persist in apomictic plants yielding new genetic variation. RAPD analysis clearly distinguished accessions and was useful in identifying sexual progenies. The implications of the novel pollen–pistil interaction on establishment and spread of apomixis in guggul are discussed. The study has not only revealed novel features of guggul reproduction but also opened new opportunities for molecular genetic analysis of sporophytic apomixis and breeding improvement of guggul.     

Detecting biodiversity hotspots using species–area and endemics–area relationships: the case of butterflies

Using data of the Red Data Book of European Butterflies we establish the species–area relationship (S = 8.5 A^0.23) and the endemics–area relationship (S = 0.5 A^0.18) of European butterflies. Applying confidence limits as tools for the identification of hotspot countries we show that in the case of butterflies hotspots of endemism and hotspots of overall species richness do not coincide. We introduce plots of residuals from species–area relationships shifted upwards by the 95% confidence limits of the intercept (α-values) as a new tool for identifying and ranking of hotspots.     

Climate–growth relationships of the dominant tree species from semi-arid savanna woodland in Ethiopia

Long-term climate–growth relationships, were examined in tree rings of four co-occurring tree species from semi-arid Acacia savanna woodlands in Ethiopia. The main purpose of the study was to prove the presence of annual tree rings, evaluate the relationship between radial growth and climate parameters, and evaluate the association of El Niño and drought years in Ethiopia. The results showed that all species studied form distinct growth boundaries, though differences in distinctiveness were revealed among the species. Tree rings of the evergreen Balanites aegyptiaca were separated by vessels surrounding a thin parenchyma band and the growth boundary of the deciduous acacias was characterized by thin parenchyma bands. The mean annual diameter increment ranged from 3.6 to 5.0 mm. Acacia senegal and Acacia seyal showed more enhanced growth than Acacia tortilis and B. aegyptiaca. High positive correlations were found between the tree-ring width chronologies and precipitation data, and all species showed similar response to external climate forcing, which supports the formation of one tree-ring per year. Strong declines in tree-ring width correlated remarkably well with past El Niño Southern Oscillation (ENSO) events and drought/famine periods in Ethiopia. Spectral analysis of the master tree-ring chronology indicated occurrences of periodic drought events, which fall within the spectral peak equivalent to 2–8 years. Our results proved the strong linkage between tree-ring chronologies and climate, which sheds light on the potential of dendrochronological studies developing in Ethiopia. The outcome of this study has important implications for paleoclimatic reconstructions and in restoration of degraded lands.