Posidonia oceanica (L.) Delile in its westernmost biogeographical limit (northwestern Alboran Sea): Meadows characterisation, phenology and flowering events

Posidonia oceanica is a Mediterranean endemic seagrass species that forms meadows covering ca. 2.5–4.5 millions of hectares, representing ca.25 % of the infralittoral and shallow circalittoral (down to 50m) bottoms of the Mediterranean. This seagrass is considered a habitat-engineer species and provides an elevated number of ecosystem services. In addition the Marine Strategy Framework Directive (MSFD, 2008/56/EC) includes seagrass like elements to evaluate the “Good Environmental Status” of the European coasts. Information about their phenological characteristic and structure of the meadows is needed for indicator estimations in order to establish their conservation status. The studied meadows are located in the westernmost limit of the P. oceanica distribution (North-western Alboran Sea) in the vecinity of the Strait of Gibraltar, an Atlantic-Mediterranean water transition area. Four sites were selected from East to West: Paraje Natural de Acantilados de Maro-Cerro Gordo (hereafter Maro), Special Area of Conservation “Calahonda” (hereafter Calahonda), Site of Community Importance Estepona (hereafter Estepona) and Punta Chullera (hereafter Chullera) where P. oceanica present their westernmost meadows. Phenological data were recorded from mid November to mid December in P. oceanica patches located at 2 – 3 m depth. At each site three types of patches (patch area <1m2, small patches; 1-2 m2, medium patches and >2 m2, large patches) were sampled. At each patch and site, 3 quadrants of 45 x 45 cm were sampled for shoot and inflorescences density measurements. In each quadrant, 10 random shoots were sampled for shoot morphology (shoot height and number of leaves). Shoot and inflorescences densities were standardized to squared meters. All the studied P. oceanica meadows develop on rocks and they present a fragmented structure with a coverage ranging between ca. 45% in Calahonda and Estepona and ca. 31% in Maro. The meadows of Chullera are reduced to a few small - medium patches with areas ranging between 0.5-1.5 m2 (Fig. 1). The meadows of Chullera and Estepona presented similar values of shoot density (ca. 752 – 662 shoots m-2, respectively) and leaf height (ca. 25 cm). Similarly, the Calahonda and Maro meadows also showed similar values of shoot density (ca. 510 – 550 shoots m-2, respectively) but displaying lower values than those of sites located closer to the Strait of Gibraltar. Regarding patch sizes and leaf height, the longest leaves (ca. 25 cm) were found in medium and large patches, but the number of leaves per shoot were higher in the small and the medium size patches (ca. 6.3 leaves per shoot). Flowering was only detected at the Calahonda meadows with maximum values of ca. 330 inflorescences m-2 (115.2 ± 98.2 inflorescences m-2, n= 9; mean ± SD) (Fig.1). Inflorescence density was not significant different among patches of different sizes. In the Alboran Sea and unlike the studied meadows, extensive beds of P. oceanica occur at the National Park of Cabo de Gata (northeastern Alboran Sea), but from east to west (Strait of Gibraltar), meadows are gradually fragmenting and their depth range decrease from 30m to 2m depth between Cabo de Gata and Chullera, respectively. Probably, the Atlantic influence and the characteristic oceanographic conditions of the Alboran Sea (i.e., higher turbidity, higher water turbulence) represent a developmental limiting factor for P. oceanica at higher depths. Similarities between the meadows located closer to Strait of Gibraltar (Chullera and Estepona) were detected as well as between those more distant (Calahonda and Maro). The first ones showed higher values of shoot densities and leaf heights than the formers, which could be relating to the higher hydrodynamic exposure found at Chullera and Estepona meadows. Regarding flowering events, sexual reproduction in P. oceanica is not common in different locations of the Mediterranean Sea. The available information seems to indicate that flowering represent an irregular event and it is related to high seawater temperature. In fact, the flowering episodes that occurred in Calahonda in November 2015, match with the warmest year ever recorded. This is the third flowering event registered in these meadows located close to the westernmost distributional limit of P. oceanica (Málaga, Alboran Sea), which could indicates that these meadows presents a healthy status. Furthermore, the absence of significant differences in relation to inflorescence density between patches of different sizes may be indicating that the fragmentation does not necessarily influence on the flowering of this seagrass species.

Embryogenesis in flowering plants: Recent approaches and prospects

The overall architectural pattern of the mature plant is established during embryogenesis. Very little is known about the molecular processes that underlie embryo morphogenesis. Last decade has, nevertheless, seen a burst of information on the subject. The synchronous somatic embryogenesis system of carrot is largely being used as the experimental system. Information on the molecular regulation of embryogenesis obtained with carrot somatic embryos as well as observations on sandalwood embryogenic system developed in our laboratory are summarized in this review. The basic experimental strategy of molecular analysis mostly relied on a comparison between genes and proteins being expressed in embryogenic and non-embryogenic cells as well as in the different stages of embryogenesis. Events such as expression of totipotency of cells and establishment of polarity which are so critical for embryo development have been characterized using the strategy, Several genes have been identified and cloned from the carrot system, These include sequences that encode certain extracellular proteins (EPs) that influence cell proliferation and embryogenesis in specific ways and sequences of the abscisic acid (ABA) inducible late embryogenesis abundant (LEA) proteins which are most abundant and differentially expressed mRNAs in somatic embryos. That LEAs are expressed in the somatic embryos of a tree flora also is evidenced from studies on sandalwood Several undescribed or novel sequences that are enhanced in embryos were identified. A sequence of this nature exists in sandalwood embryos was demonstrated using a Cuscuta haustorial (organ-specific) cDNA probe. Somatic embryogenesis systems have been used to assess the expression of genes isolated from non-embryogenic tissues. Particular attention has been focused on both cell cycle and histone genes.

Flowering in British Lemna: A rare, cyclic or simply overlooked phenomenon?

It is largely presumed that reproduction in British Lemna, as in other British Lemnaceae, is almost entirely asexual, with new daughter fronds being produced from the side pouches of older mother fronds. Sexual reproduction is considered to be a rather rare event or even absent and because of this rarity the sexual features of Lemna, such as anthers and fruit, are often considered to be of little taxonomic value. It was with some surprise, therefore, that widespread flowering was observed in all British Lemna during the summer of 1995. Initial observations in Shropshire during June recorded flowers in minor and trisulca, with fruit production in trisulca. L.gibba, minor and minuta were noted as being in flower on several occasions in Kent, during July and August, probably fruit production occurring in both species. To what extent these events are truly representative of the sexual reproduction rate of British Lemna on a year-to-year basis, or simply reflect the unusually high summer temperatures of 1995, is unclear.

Temperate flowering phenology

Individuals, families, networks, and botanic gardens have made records of flowering times of a wide range of plant species over many years. These data can highlight year to year changes in seasonal events (phenology) and those datasets covering long periods draw interest for their perspective on plant responses to climate change. Temperate flowering phenology is complex, using environmental cues such as temperature and photoperiod to attune flowering to appropriate seasonal conditions. Here we give an overview of flowering phenological recording, outline different patterns of flowering, and look at the interpretation of datasets in relation to seasonal and climatic change.

Asynchronous flowering and within-plant flowering diversity in wheat and the implications for crop resilience to heat

Self-pollination dominates in wheat , with a small level of out-crossing due to flowering asynchrony and male sterility. However, the timing and synchrony of male and female flowering in wheat is a crucial determinant of seed set and may be an important factor affecting gene flow and resilience to climate change. Here, a methodology is presented for assessing the timing and synchrony of flowering in wheat. From the onset of flowering until the end of anthesis, the anther and stigma activity of each floret was assessed on the first five developing ears in potted plants grown under ambient conditions and originating from cv Paragon or cvs Spark-Rialto backgrounds. At harvest maturity, seed presence, size and weight was recorded for each floret scored. The synchrony between pollen dehiscence and stigma collapse within a flower was dependent on its relative position in a spike and within a floret. Determined on the basis of synchrony within each flower, the level of pollination by pollen originating from other flowers reached approximately 30% and did not change throughout the duration of flowering. A modelling exercise parameterised by flowering observations indicated that the temporal and spatial variability of anther activity within and between spikes may influence the relative resilience of wheat to sudden, extreme climatic events which has direct relevance to predicted future climate scenarios in the UK.

Genome dynamics explain the evolution of flowering time CCT domain gene families in the Poaceae

Numerous CCT domain genes are known to control flowering in plants. They belong to the CONSTANS-like (COL) and PREUDORESPONSE REGULATOR (PRR) gene families, which in addition to a CCT domain possess B-box or response-regulator domains, respectively. Ghd7 is the most recently identified COL gene to have a proven role in the control of flowering time in the Poaceae. However, as it lacks B-box domains, its inclusion within the COL gene family, technically, is incorrect. Here, we show Ghd7 belongs to a larger family of previously uncharacterized Poaceae genes which possess just a single CCT domain, termed here CCT MOTIF FAMILY (CMF) genes. We molecularly describe the CMF (and related COL and PRR) gene families in four sequenced Poaceae species, as well as in the draft genome assembly of barley (Hordeum vulgare). Genetic mapping of the ten barley CMF genes identified, as well as twelve previously unmapped HvCOL and HvPRR genes, finds the majority map to colinear positions relative to their Poaceae orthologues. Combined inter-/intra-species comparative and phylogenetic analysis of CMF, COL and PRR gene families indicates they evolved prior to the monocot/dicot divergence ~200 mya, with Poaceae CMF evolution described as the interplay between whole genome duplication in the ancestral cereal, and subsequent clade-specific mutation, deletion and duplication events. Given the proven role of CMF genes in the modulation of cereals flowering, the molecular, phylogenetic and comparative analysis of the Poaceae CMF, COL and PRR gene families presented here provides the foundation from which functional investigation can be undertaken.

Genomic analyses of flowering transition and inflorescence development in grapevine = Análisis genómicos de la transición floral y del desarrollo de la inflorescencia en la vid

The general objective of this work is to analyze the regulatory processes underlying flowering transition and inflorescence and flower development in grapevine. Most of these crucial developmental events take place within buds growing during two seasons in two consecutive years. During the first season, the shoot apical meristem within the bud differentiates all the basic elements of the shoot including flowering transition in lateral primordia and development of inflorescence primordia. These events practically end with bud dormancy. The second season, buds resume shoot growth associated to flower formation and development. In grapevine, the lateral meristems can give rise either to tendril or inflorescence primordia that are homologous organs. With this purpose, we performed global transcriptome analyses along the bud annual cycle and during inflorescence and tendril development. In addition, we approach the genomic analysis of the MIKC type MADS-box gene family in grapevine to identify all its members and assign them putative biological functions. Regarding buds developmental cycle, the results indicate that the main factors explaining the global gene expression differences were the processes of bud dormancy and active growth as well as stress responses. Non dormant buds exhibited up-regulation in functional categories typical of actively proliferating and growing cells (photosynthesis, cell cycle regulation, chromatin assembly) whereas in dormant ones the main functional categories up-regulated were associated to stress response pathways together with transcripts related to starch catabolism. Major transcriptional changes during the dormancy period were associated to the para/endodormancy, endo/ecodormancy and ecodormancy/bud break transitions. Global transcriptional analyses along tendril and inflorescence development suggested that these two homologous organs share a common transcriptional program related to cell proliferation functions. Both structures showed a progressive decrease in the expression of categories such as cell-cycle, auxin metabolism/signaling, DNA metabolism, chromatin assembly and a cluster of five transcripts belonging to the GROWTH-REGULATING FACTOR (GRF) transcription factor family, that are known to control cell proliferation in other species and determine the size of lateral organs. However, they also showed organ specific transcriptional programs that can be related to their differential organ structure and function. Tendrils showed higher transcription of genes related to photosynthesis, hormone signaling and secondary metabolism than inflorescences, while inflorescences have higher transcriptional activity for genes encoding transcription factors (especially those belonging to the MADS-box gene family). Further analysis along inflorescence development evidenced the relevance of additional functions likely related to processes of flower development such as fatty acid and lipid metabolism, jasmonate signaling and oxylipin biosynthesis. The transcriptional analyses performed highlighted the relevance of several groups of transcriptional regulators in the developmental processes studied. The expression profiles along bud development revealed significant differences for some MADS-box subfamilies in relation to other plant species, like the members of the FLC and SVP subfamilies suggesting new roles for these groups in grapevine. In this way, it was found that VvFLC2 and VvAGL15.1 could participate, together with some members of the SPL-L family, in dormancy regulation, as was shown for some of them in other woody plants. Similarly, the expression patterns of the VvFLC1, VvFUL, VvSOC1.1 (together with VvFT, VvMFT1 and VFL) genes could indicate that they play a role in flowering transition in grapevine, in parallel to their roles in other plant systems. The expression levels of VFL, the grapevine LEAFY homolog, could be crucial to specify the development of inflorescence and flower meristems instead of tendril meristems. MADS-box genes VvAP3.1 and 2, VvPI, VvAG1 and 3, VvSEP1-4, as well as VvBS1 and 2 are likely associated with the events of flower meristems and flower organs differentiation, while VvAP1 and VvFUL-L (together with VvSOC1.1, VvAGL6.2) could be involved on tendril development given their expression patterns. In addition, the biological function ofVvAP1 and VvTFL1A was analyzed using a gene silencing approach in transgenic grapevine plants. Our preliminary results suggested a possible role for both genes in the initiation and differentiation of tendrils. Finally, the genomic analysis of the MADS-box gene family in grapevine revealed differential features regarding number and expression pattern of genes putatively involved in the flowering transition process as compared to those involved in the specification of flower and fruit organ identity. Altogether, the results obtained allow identifying putative candidate genes and pathways regulating grapevine reproductive developmental processes paving the way to future experiments demonstrating specific gene biological functions. RESUMEN El objetivo general de este trabajo es analizar los procesos regulatorios subyacentes a la inducción floral así como al desarrollo de la inflorescencia y la flor en la vid. La mayor parte de estos eventos cruciales tienen lugar en las yemas a lo largo de dos estaciones de crecimiento consecutivas. Durante la primera estación, el meristemo apical contenido en la yema diferencia los elementos básicos del pámpano, lo cual incluye la inducción de la floración en los meristemos laterales y el subsiguiente desarrollo de primordios de inflorescencia. Estos procesos prácticamente cesan con la entrada en dormición de la yema. En la segunda estación, se reanuda el crecimiento del pámpano acompañado por la formación y desarrollo de las flores. En la vid, los meristemos laterales pueden dar lugar a primordios de inflorescencia o de zarcillo que son considerados órganos homólogos. Con este objetivo llevamos a cabo un estudio a nivel del transcriptoma de la yema a lo largo de su ciclo anual, así como a lo largo del desarrollo de la inflorescencia y del zarcillo. Además realizamos un análisis genómico de la familia MADS de factores transcripcionales (concretamente aquellos del tipo MIKC) para identificar todos sus miembros y tratar de asignarles posibles funciones biológicas. En cuanto al ciclo de desarrollo de la yema, los resultados indican que los principales factores que explican las diferencias globales en la expresión génica fueron los procesos de dormición de la yema y el crecimiento activo junto con las respuestas a diversos tipos de estrés. Las yemas no durmientes mostraron un incremento en la expresión de genes contenidos en categorías funcionales típicas de células en proliferación y crecimiento activo (como fotosíntesis, regulación del ciclo celular, ensamblaje de cromatina), mientras que en las yemas durmientes, las principales categorías funcionales activadas estaban asociadas a respuestas a estrés, así como con el catabolismo de almidón. Los mayores cambios observados a nivel de transcriptoma en la yema coincidieron con las transiciones de para/endodormición, endo/ecodormición y ecodormición/brotación. Los análisis transcripcionales globales a lo largo del desarrollo del zarcillo y de la inflorescencia sugirieron que estos dos órganos homólogos comparten un programa transcripcional común, relacionado con funciones de proliferación celular. Ambas estructuras mostraron un descenso progresivo en la expresión de genes pertenecientes a categorías funcionales como regulación del ciclo celular, metabolismo/señalización por auxinas, metabolismo de ADN, ensamblaje de cromatina y un grupo de cinco tránscritos pertenecientes a la familia de factores transcripcionales GROWTH-REGULATING FACTOR (GRF), que han sido asociados con el control de la proliferación celular y en determinar el tamaño de los órganos laterales en otras especies. Sin embargo, también pusieron de manifiesto programas transcripcionales que podrían estar relacionados con la diferente estructura y función de dichos órganos. Los zarcillos mostraron mayor actividad transcripcional de genes relacionados con fotosíntesis, señalización hormonal y metabolismo secundario que las inflorescencias, mientras que éstas presentaron mayor actividad transcripcional de genes codificantes de factores de transcripción (especialmente los pertenecientes a la familia MADS-box). Análisis adicionales a lo largo del desarrollo de la inflorescencia evidenciaron la relevancia de otras funciones posiblemente relacionadas con el desarrollo floral, como el metabolismo de lípidos y ácidos grasos, la señalización mediada por jasmonato y la biosíntesis de oxilipinas. Los análisis transcripcionales llevados a cabo pusieron de manifiesto la relevancia de varios grupos de factores transcripcionales en los procesos estudiados. Los perfiles de expresión estudiados a lo largo del desarrollo de la yema mostraron diferencias significativas en algunas de las subfamilias de genes MADS con respecto a otras especies vegetales, como las observadas en los miembros de las subfamilias FLC y SVP, lo cual sugiere que podrían desempeñar nuevas funciones en la vid. En este sentido, se encontró que los genes VvFLC2 y VvAGL15.1 podrían participar, junto con algunos miembros de la familia SPL-L, en la regulación de la dormición. De un modo similar, los patrones de expresión de los genes VvFLC1, VvFUL, VvSOC1.1 (junto con VvFT, VvMFT1 y VFL) podría indicar que desempeñan un papel en la regulación de la inducción de la floración en la vid, como se ha observado en otros sistemas vegetales. Los niveles de expresión de VFL, el homólogo en vid del gen LEAFY de A. thaliana podrían ser cruciales para la especificación del desarrollo de meristemos de inflorescencia y flor en lugar de meristemos de zarcillo. Los genes VvAP3.1 y 2, VvPI, VvAG1 y 3, VvSEP1-4, así como VvBS1 y 2 parecen estar asociados con los eventos de diferenciación de meristemos y órganos florales, mientras que VvAP1 y VvFUL-L (junto con VvSOC1.1 y VvAGL6.2) podrían estar implicados en el desarrollo del zarcillo dados sus patrones de expresión. Adicionalmente, se analizó la función biológica de los genes VvAP1 y VvTFL1A por medio de una estrategia de silenciamiento génico. Los datos preliminares sugieren un posible papel para ambos genes en la iniciación y diferenciación de los zarcillos. Finalmente, el análisis genómico de la familia MADS en vid evidenció diferencias con respecto a otras especies vegetales en cuanto a número de miembros y patrón de expresión en genes supuestamente implicados en la inducción de la floración, en comparación con aquellos relacionados con la especificación de identidad de órganos florales y desarrollo del fruto. En conjunto, los resultados obtenidos han permitido identificar posibles rutas y genes candidatos a participar en la regulación de los procesos de desarrollo reproductivo de la vid, sentando las bases de futuros experimentos encaminados a conocer la funciones biológicas de genes específicos.

A calcium influx is triggered and propagates in the zygote as a wavefront during in vitro fertilization of flowering plants

In this paper, we report direct measurement of an influx of extracellular Ca2+ induced by gamete fusion in flowering plants. This result was obtained during maize in vitro fertilization with the use of an extracellular Ca2+-selective vibrating probe. Ca2+ influx recorded at the surface of isolated egg cells, with or without adhesion of a male sperm cell, was close to zero and stable over time. Gamete fusion, however, triggered a Ca2+ influx in the vicinity of the sperm entry site with a delay of 1.8 ± 0.6 sec. The Ca2+ influx spread subsequently through the whole egg cell plasma membrane as a wavefront, progressing at an estimated rate of 1.13 μm⋅sec−1. Once established, Ca2+ influx intensities were sustained, monotonic and homogeneous over the whole egg cell, with an average peak influx of 14.92 pmol⋅cm−2⋅sec−1 and an average duration of 24.4 min. The wavefront spread of channel activation correlates well with the cytological modifications induced by fertilization, such as egg cell contraction, and with the cytosolic Ca2+ (c[Ca2+]) elevation previously reported. Calcium influx was inhibited effectively by gadolinium, possibly implicating mechanosensitive channels. Furthermore, artificial influxes created by incubation with Ca2+ ionophores mimicked some aspects of egg activation. Taken together, these results suggest that, during fertilization in higher plants, gamete membrane fusion starts the first embryonic events by channel opening and Ca2+ influx. In turn, c[Ca2+] may work as a trigger and possibly a space and time coordinator of many aspects of egg activation.

Adventitious root formation in cuttings of Backhousia citriodora F. Muell 2. Seasonal influences of temperature, rainfall, flowering and auxins on the stock plant

A 2-year study was carried out on established trees at two sites in southeastern Queensland, Australia, to identify environmental factors that influenced rooting of Backhousia citriodora from cuttings. Complex interactions of rainfall events above 20 mm from the preceding month and mean maximum temperature on stock plants resulted in a correlation with rooting success of r = 0.81 and 0.74 for sites at The University Of Queensland, Gatton Campus, and Cedar Glen, respectively. A more detailed investigation under controlled environmental conditions showed that maintaining stock plants at temperatures between 10 and 30degreesC had no direct effect on rooting capacity. However, temperature was correlated with growth, which may have an indirect effect on root formation. In spring floral initiation was found to only delay rooting and had no effect on the final rooting percentage. A series of seasonal experiments demonstrated that application of the auxins indole-3-acetic acid, indole-3-butyric acid and napthaleneacetic acid over a range of concentrations (1000-8000 mug/ml) did not significantly increase rooting compared to the control and there is no practical advantage in applying auxins. Seasonal results and the temperature experiment also suggest that under a glasshouse environment with higher temperatures in winter and an adequate supply of water, B. citriodora cuttings can be successfully rooted over the whole year. (C) 2004 Elsevier B.V. All rights reserved.

An investigation of nucleation events in a coastal urban environment in the southern hemisphere

The occurrence of and conditions favourable to nucleation were investigated at an industrial and commercial coastal location in Brisbane, Australia during five different campaigns covering a total period of 13 months. To identify potential nucleation events, the difference in number concentration in the size range 14-30 nm (N14-30) between consecutive observations was calculated using first-order differencing. The data showed that nucleation events were a rare occurrence, and that in the absence of nucleation the particle number was dominated by particles in the range 30-300 nm. In many instances, total particle concentration declined during nucleation. There was no clear pattern in change in NO and NO2 concentrations during the events. SO2 concentration, in the majority of cases, declined during nucleation but there were exceptions. Most events took place in summer, followed by winter and then spring, and no events were observed for the autumn campaigns. The events were associated with sea breeze and long-range transport. Roadside emissions, in contrast, did not contribute to nucleation, probably due to the predominance of particles in the range 50-100 nm associated with these emissions.