Global and regional drivers of abundance patterns in the hart's tongue fern complex (Aspleniaceae).

BACKGROUND AND AIMS: The hart's tongue fern (HTF) complex is a monophyletic group composed of five geographically segregated members with divergent abundance patterns across its broad geographic range. We postulated hierarchical systems of environmental controls in which climatic and land-use change drive abundance patterns at the global scale, while various ecological conditions function as finer scale determinants that further increase geographic disparities at regional to local scales. METHODS: After quantifying the abundance patterns of the HTF complex, we estimated their correlations with global climate and land-use dynamics. Regional determinants were assessed using boosted regression tree models with 18 potential ecological variables. Moreover, we investigated long-term population trends in the USA to understand the interplay of climate change and anthropogenic activities on a temporal scale. KEY RESULTS: Latitudinal climate shifts drove latitudinal abundance gradients, and regionally different levels of land-use change resulted in global geographic disparities in population abundance. At a regional scale, population isolation, which accounts for rescue effects, played an important role, particularly in Europe and East Asia where several hot spots occurred. Furthermore, the variables most strongly influencing abundance patterns greatly differed by region: precipitation seasonality in Europe; spatial heterogeneity of temperature and precipitation in East Asia; and magnitudes of past climate change, temperature seasonality and edaphic conditions in North America. In the USA, protected populations showed increasing trends compared with unprotected populations at the same latitude, highlighting the critical role of habitat protection in conservation measures. CONCLUSIONS: Geographic disparities in the abundance patterns of the HTF complex were determined by hierarchical systems of environmental controls, wherein climatic and land-use dynamics act globally but are modulated by various regional and local determinants operating at increasingly finer scales. We highlighted that fern conservation must be tailored to particular geographic contexts and environmental conditions by incorporating a better understanding of the dynamics acting at different spatiotemporal scales.

The complete chloroplast genome sequence of Asplenium komarovii Akasawa, a rare fern in South Korea.

We sequenced the complete chloroplast genome of Asplenium komarovii Akasawa (syn: Asplenium scolopendrium L. subsp. japonicum (Komarov) Rasbach, Reichstein & Viane), which is designated as a rare species in South Korea. The complete chloroplast genome is 149,393 bp in total length and comprised of the following regions: large single copy (82,464 bp), small single copy (21,345 bp), and a pair of inverted repeats (22,792 bp). The overall GC content is 40.9% and the genome encoded a total of 115 genes, including 84 protein-coding, 27 transfer RNA, and 4 ribosomal RNA genes. Phylogenetic analysis based on 21 representative chloroplast genomes of the suborder Aspleniineae (and one outgroup) indicates that Aspleniaceae is monophyletic and sister to Diplaziopsidaceae, with Rhadidosoraceae as the basal group in this three family clade. Asplenium komarovii is sister to A. nidus and A. prolongatum with strong bootstrap support. The chloroplast genome of A. komarovii will be useful in establishing its relationships within the A. scolopendrium complex, which is currently unresolved.

Expression patterns of conserved microRNAs in the male gametophyte of loblolly pine (Pinus taeda).

MicroRNAs (miRNAs) are small RNAs that regulate genes involved in various aspects of plant development, but their presence and expression patterns in the male gametophytes of gymnosperms have not yet been established. Therefore, this study identified and compared the expression patterns of conserved miRNAs from two stages of the male gametophyte of loblolly pine (Pinus taeda), which are the mature (ungerminated) and germinated pollen. Microarray was used to identify conserved miRNAs that varied in expression between these two stages of the loblolly pine male gametophyte. Forty-seven conserved miRNAs showed significantly different expression levels between mature and germinated loblolly pine pollen. In particular, miRNAs representing 14 and 8 families were up- and down-regulated in germinated loblolly pine pollen, respectively. qRT-PCR was used to validate their expression patterns using representative miRNAs. Target genes and proteins were identified using psRNATarget program. Predicted targets of the 22 miRNA families belong mostly to classes of genes involved in defense/stress response, metabolism, regulation, and signaling. qRT-PCR was also used to validate the expression patterns of representative target genes. This study shows that conserved miRNAs are expressed in mature and germinated loblolly pine pollen. Many of these miRNAs are differentially expressed, which indicates that the two stages of the male gametophyte examined are regulated at the miRNA level. This study also expands our knowledge of the male gametophytes of seed plants by providing insights on some similarities and differences in the types and expression patterns of conserved miRNAs between loblolly pine with those of rice and Arabidopsis.

Anatomical aspects of angiosperm root evolution.

BACKGROUND AND AIMS: Anatomy had been one of the foundations in our understanding of plant evolutionary trends and, although recent evo-devo concepts are mostly based on molecular genetics, classical structural information remains useful as ever. Of the various plant organs, the roots have been the least studied, primarily because of the difficulty in obtaining materials, particularly from large woody species. Therefore, this review aims to provide an overview of the information that has accumulated on the anatomy of angiosperm roots and to present possible evolutionary trends between representatives of the major angiosperm clades. SCOPE: This review covers an overview of the various aspects of the evolutionary origin of the root. The results and discussion focus on angiosperm root anatomy and evolution covering representatives from basal angiosperms, magnoliids, monocots and eudicots. We use information from the literature as well as new data from our own research. KEY FINDINGS: The organization of the root apical meristem (RAM) of Nymphaeales allows for the ground meristem and protoderm to be derived from the same group of initials, similar to those of the monocots, whereas in Amborellales, magnoliids and eudicots, it is their protoderm and lateral rootcap which are derived from the same group of initials. Most members of Nymphaeales are similar to monocots in having ephemeral primary roots and so adventitious roots predominate, whereas Amborellales, Austrobaileyales, magnoliids and eudicots are generally characterized by having primary roots that give rise to a taproot system. Nymphaeales and monocots often have polyarch (heptarch or more) steles, whereas the rest of the basal angiosperms, magnoliids and eudicots usually have diarch to hexarch steles. CONCLUSIONS: Angiosperms exhibit highly varied structural patterns in RAM organization; cortex, epidermis and rootcap origins; and stele patterns. Generally, however, Amborellales, magnoliids and, possibly, Austrobaileyales are more similar to eudicots, and the Nymphaeales are strongly structurally associated with the monocots, especially the Acorales.

Constitutive expression of the SAP1 gene from willow (Salix discolor) causes early flowering in Arabidopsis thaliana.

SAP1-1 and SAP1-2 were isolated from the male reproductive buds of willow (Salix discolor, clone S365). SAP1-1 differs from SAP1-2 based on a few nucleotide substitutions, but the sizes of their full-length cDNAs are identical. The deduced amino acid sequences of SAP1-1 and SAP1-2 were 98% similar and contain the same C-terminal amino acid motif "GYGA" like that of PTAP1-2 from Populus trichocarpa. The expression patterns of SAP1 in various parts of the male reproductive buds of S. discolor implicate this gene in the formation of the inflorescence meristems, bracts, and floral meristems. To characterize the functions of SAP1, we assessed Arabidopsis thaliana transformed with 35S: :SAP1-1. A total of 52 transgenic T1 lines were obtained, and a 3:1 segregation ratio was obtained in the T2 generation of each line. In the T3 generation, five homozygous transgenic lines were obtained, which were used for further analysis. Screening of transgenic lines was greatly facilitated by the detection of GFP expression starting with germinating seeds. Phenotypes of the homozygous transgenic lines included early flowering, conversion of inflorescence branches to solitary flowers, formation of terminal flowers, and formation of flowers with greater number of petals, stamens, and pistils. Northern analysis showed similar expression levels in all five lines. This study provides the first functional analysis of an APETALA1 (AP1)/SQUAMOSA (SQUA) homolog from a dioecious species and suggests that SAP1 is a homolog of the AP1/SQUA gene.

In vitro germination and transient GFP expression of American chestnut (Castanea dentata) pollen.

The development of the male reproductive structures of American chestnut (Castanea dentata) is described to advance our understanding of its reproductive behavior. This information has been vital in the development of a strategy to collect pollen grains from male catkins suitable for in vitro germination and transformation experiments. Cutting male catkins into small segments and rolling them over a culture plate resulted in evenly dispersed and large amounts of pollen with minimal unwanted accessory floral parts. To optimize pollen viability, the effect of various storage conditions on in vitro germination was examined. Our results showed that initial storage at 4 degrees C for 2 weeks significantly increased percent germination as compared to freshly collected pollen and those stored directly at -20 degrees C or -80 degrees C. This also means that for long-term storage of American chestnut pollen, the catkins should first be kept at 4 degrees C for a couple of weeks and then at -80 degrees C. The use of pollen grains with high viability is necessary for the transformation of American chestnut pollen. To optimize pollen transformation via particle bombardment, the effects of target distance, target pressure, and pollen developmental stage were examined. Statistical analysis showed that bombardment of ungerminated pollen at 1,100 psi resulted in the highest percent transient GFP expression (4.1%).

Characterization of pollen tube development in Pinus strobus (Eastern white pine) through proteomic analysis of differentially expressed proteins.

The differentially expressed proteins in pollen tubes indicate their specific roles in this stage of male gametophyte development. To isolate these proteins, 2-DE was done using ungerminated pollen and 2-day-old pollen tubes of Pinus strobus. Results show that 645 and 647 protein spots were clearly resolved from pollen grains and pollen tubes, respectively. Thirty-eight protein spots were expressed only in pollen tubes, while 19 increased in intensity. MALDI-TOF MS was used to generate tryptic peptide masses that were submitted to Mascot for identification. Of the differentially expressed proteins, 12% matched with hypothetical proteins, 33% did not hit any protein, and for the 55%, a putative function was assigned based on similarity of sequences with previously characterized proteins. Therefore, pollen tube development can be characterized by the cellular activities that involve metabolism, stress/defense response, gene regulation, signal transduction, and cell wall formation. This study expands our understanding of the changes in protein expression associated with pollen tube development and provides insights into the molecular programs that separate the development of the pollen tubes from pollen grains. This is the first report that describes a global analysis of differentially expressed proteins from the pollen tube of any seed plant.