DNA amounts for five pteridophyte species fill phylogenetic gaps in C-value data

C-values are important biodiversity characters, yet data for pteridophytes are scarce. A recent survey showed that reliable C-value data were available for only 62 species. This paper reports C-value estimates for a further five pteridophyte species which were selected to fill phylogenetic gaps identified in previous work: Angiopteris lygodiifolia , of the eusporangiate fern family Marattiaceae (1C = 7.10 pg); Lygodium japonicum , of the basal leptosporangiate fern family Schizaeaceae (1C = 11.66 pg); Marsilea quadrifolia , of the heterosporous water fern order Marsileales (1C = 4.01 pg); Isoetes lacustris (1C = 11.97 pg) and Lycopodium clavatum (1C = 2.86 pg), of the lycophyte families Isoetaceae and Lycopodiaceae, respectively. The C-value estimates are discussed in the context of other C-value data. Further, the range of C-values in lycophytes (0.16 pg – 11.97 pg), which comprise Selaginellaceae, Isoetaceae and Lycopodiaceae, is discussed in relation to nucleotypic constraints imposed by sperm flagella number.  © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 140 , 169−173.     

Genome size evolution of the extant lycophytes and ferns

Ferns and lycophytes have remarkably large genomes. However, little is known about how their genome size evolved in fern lineages. To explore the origins and evolution of chromosome numbers and genome size in ferns, we used flow cytometry to measure the genomes of 240 species (255 samples) of extant ferns and lycophytes comprising 27 families and 72 genera, of which 228 species (242 samples) represent new reports. We analyzed correlations among genome size, spore size, chromosomal features, phylogeny, and habitat type preference within a phylogenetic framework. We also applied ANOVA and multinomial logistic regression analysis to preference of habitat type and genome size. Using the phylogeny, we conducted ancestral character reconstruction for habitat types and tested whether genome size changes simultaneously with shifts in habitat preference. We found that 2C values had weak phylogenetic signal, whereas the base number of chromosomes (x) had a strong phylogenetic signal. Furthermore, our analyses revealed a positive correlation between genome size and chromosome traits, indicating that the base number of chromosomes (x), chromosome size, and polyploidization may be primary contributors to genome expansion in ferns and lycophytes. Genome sizes in different habitat types varied significantly and were significantly correlated with habitat types; specifically, multinomial logistic regression indicated that species with larger 2C values were more likely to be epiphytes. Terrestrial habitat is inferred to be ancestral for both extant ferns and lycophytes, whereas transitions to other habitat types occurred as the major clades emerged. Shifts in habitat types appear be followed by periods of genomic stability. Based on these results, we inferred that habitat type changes and multiple whole-genome duplications have contributed to the formation of large genomes of ferns and their allies during their evolutionary history.     

Intercontinental disjunctions between eastern Asia and western North America in vascular plants highlight the biogeographic importance of the Bering land bridge from late Cretaceous to Neogene

This review shows a close biogeographic connection between eastern Asia and western North America from the late Cretaceous to the late Neogene in major lineages of vascular plants (flowering plants, gymnosperms, ferns and lycophytes). Of the eastern Asian–North American disjuncts, conifers exhibit a high proportion of disjuncts between eastern Asia and western North America. Several lineages of ferns also show a recent disjunct pattern in the two areas. In flowering plants, the pattern is commonly shown in temperate elements between northeastern Asia and northwestern North America, as well as elements of the relict boreotropical and Neogene mesophytic and coniferous floras. The many cases of intercontinental biogeographic disjunctions between eastern Asia and western North America in plants supported by recent phylogenetic analyses highlight the importance of the Bering land bridge and/or the plant migrations across the Beringian region from the late Cretaceous to the late Neogene, especially during the Miocene. The Beringian region has permitted the filtering and migration of certain plant taxa since the Pliocene after the opening of the Bering Strait, as many conspecific taxa or closely related species occur on both sides of Beringia.     

Molecular Systematic Study of <Emphasis Type="Italic">Chrysosplenium</Emphasis> Series <Emphasis Type="Italic">Pilosa</Emphasis> (Saxifragaceae) in Korea

To address systematic issues pertaining to series Pilosa of Korean Chrysosplenium, the phylogenetic relationships of the 35 accessions of Korean Chrysosplenium taxa were examined using the DNA sequence data obtained from the nuclear ribosomal internal transcribed spacer (ITS) regions and the psbK-I gene of the plastid genome. The results from both ITS and psbK-I sequence data indicated that Chrysosplenium flaviflorum formed a well-supported clade that merits species status of the taxon. The ITS phylogenetic trees detected two distinct lineages in Chrysosplenium pilosum var. fulvum. Geographically, one lineage was widely distributed throughout the country whereas the other was confined to Gyeonggi and Gangwon provinces. The plants sampled from Jeju Island, which have been recognized as C. pilosum var. sphaerospermum [C. hallaisanense] exhibited no DNA sequence feature segregating them from C. pilosum var. fulvum [C. barbatum] distributed in the mainland area of Korea. The plastid DNA sequence data was not fully in concordance with the nuclear ribosomal ITS data and the current taxa delimitation. These results suggest incomplete lineage sorting or that plastid genome capture may have occurred during the evolution of some taxa examined in the present study.     

Construction of a bacterial artificial chromosome library from the spikemoss Selaginella moellendorffii: a new resource for plant comparative genomics

Background

The lycophytes are an ancient lineage of vascular plants that diverged from the seed plant lineage about 400 Myr ago. Although the lycophytes occupy an important phylogenetic position for understanding the evolution of plants and their genomes, no genomic resources exist for this group of plants.

Results

Here we describe the construction of a large-insert bacterial artificial chromosome (BAC) library from the lycophyte Selaginella moellendorffii. Based on cell flow cytometry, this species has the smallest genome size among the different lycophytes tested, including Huperzia lucidula, Diphaiastrum digita, Isoetes engelmanii and S. kraussiana. The arrayed BAC library consists of 9126 clones; the average insert size is estimated to be 122 kb. Inserts of chloroplast origin account for 2.3% of the clones. The BAC library contains an estimated ten genome-equivalents based on DNA hybridizations using five single-copy and two duplicated S. moellendorffii genes as probes.

Conclusion

The S. moellenforffii BAC library, the first to be constructed from a lycophyte, will be useful to the scientific community as a resource for comparative plant genomics and evolution.     

Assessment of phylogenetic inter-relationships in the genus Cymbidium (Orchidaceae) based on internal transcribed spacer region of rDNA

Sequence data obtained from nrITS region were used to assess phylogenetic inter-relationships and infrageneric classification of ten Cymbidium species collected from north-east India. The final aligned data matrix of combined ITS 1, 5.8S and ITS 2 yielded 684 characters. The ITS 1 and ITS 2 regions showed variable sequence lengths and G + C content (%). The 5.8S region was found to be more conserved (98.71%) followed by ITS 1 (86.12%) and ITS 2 (69.40%). ITS 2 recorded highest percentage of parsimony informative sites (7.46%), high sequence divergence with indels (24.63%), high number of transitions and transversions. ITS sequence data determined the phylogeny of Asiatic Cymbidiums with high bootstrap values. All three proposed subgenera could be distinguished clearly by all four (MP, ML, NJ, and BI) phylogenetic methods. This study validates the utility of ITS rDNA region as a reliable indicator of phylogenetic relationships, especially ITS 2 as probable DNA barcode at higher levels and can serve as an additional approach for identification of broader range of plant taxa especially orchids.     

中国石松类和蕨类植物多样性研究进展

石松类和蕨类植物是维管植物的第二大类群, 其起源可追溯到4亿年前。在被子植物出现之前, 石松类和蕨类植物在古地球生态系统中占主导地位, 其重要性一直延续到现在。自20世纪40年代开始, 中国石松类和蕨类植物研究就令世界瞩目, 尤其是2017年第19届国际植物学大会在中国深圳召开后的5年时间里, 中国石松类和蕨类植物研究更是面向世界、走向国际, 研究更为广泛的科学问题, 在物种多样性、保护、系统演化和生态适应性等方面取得了一系列重要研究进展。2017-2022年, 多个中国研究团队利用多组学数据构建了世界石松类和蕨类植物科级水平的生命之树并提出了关键性状孢子囊环带演化的新模式; 解决了石松类和蕨类植物中目级、科级、属级和种级众多关键的系统分类学等问题, 发表了106个新分类群; 开展了大量的植物区系调查和研究, 出版了6部中国石松类和蕨类植物多样性专著和1部世界性专著; 对65种国家重点保护的石松类和蕨类植物进行了迁地保护, 同时实现了桫椤科、水蕨属(Ceratopteris)、观音座莲属(Angiopteris)和鹿角蕨(Platycerium wallichii)等重点保护类群的孢子繁殖; 在系统发育框架下, 研究了石松类和蕨类植物的生态修复功能和生态适应性演化。通过对2017-2022年研究成果的总结和思考, 本文对未来石松类和蕨类植物的发展提出以下建议: (1)提高中国寡型科属以及世界性大科大属的关注力度; (2)加强西藏、四川等薄弱地区石松类和蕨类植物的调查研究, 并结合新技术, 如DNA条形码等以提高区系调查中物种鉴别的效率和准确性; (3)运用多学科交叉的研究方法厘清各科、属、种间系统关系的同时, 还应加强系统和生态适应性演化之间的协同研究; (4)关注石松类和蕨类植物系统位置作为陆生维管植物演化起点的共性科学问题; (5)加强石松类和蕨类植物系统分类学与生态学、植物化学、保护生物学等学科间交叉合作研究。     

The Exploitation and Utilization of Lycophyte and Fern Resources in Xishuangbanna,Yunnan

Xishuangbanna lies in the south of Yunnan. The composition and distributive regulation of lycophytes and ferns in Xishuangbanna are studied in this paper. As a result, 363 species of wild lycophytes and ferns which belong to 76 genera and 31 families are found in this area. The six species rich families are Polypodiaceae, Athyriaceae, Thelypteridaceae, Aspleniaceae, Pteridaceae and Selaginellaceae. The geographical distribution of the species shows that this flora mainly consists of Tropical Asia elements. Terrestrial species are the richest of this flora, followed by epiphytic and lithophytic species. The lycophytes and ferns resources of Xishuangbanna are abundant. However, the exploitation and utilization of this group of plants are ignored. This study investigated lycophytes and ferns resources of Xishuangbanna. Medicinal, edible and ornamental ferns in Xishuangbanna have been clearly researched and their utilization and protection have been discussed.     

Evolution of shoot apical meristem structures in vascular plants with respect to plasmodesmatal network

Vascular plants have evolved shoot apical meristems (SAMs), whose structures differ among plant groups. To clarify the evolutionary course of the different structural types of SAMs, we compared plasmodesmatal networks in the SAMs for 17 families and 24 species of angiosperms, gymnosperms, and pteridophytes, using transmission electron microscopy (TEM). The plasmodesmata (PD) in almost all cell walls in median longitudinal sections of SAMs were counted, and the PD density per unit area was calculated for each cell wall. Angiosperm and gymnosperm SAMs have low densities, with no difference between stratified (tunica-corpus) and unstratified structures. SAMs of ferns, including Psilotum and Equisetum, have average densities that are more than three times higher than those of seed plants. Interestingly, microphyllous lycopods have both the fern and seed-plant types of PD networks; Selaginellaceae SAMs with single apical cells have high PD densities, while SAMs of Lycopodiaceae and Isoetaceae with plural initial cells have low PD densities, equivalent to those of seed plants. In summary, PD networks are strongly correlated to SAM organizations-SAMs with single and plural initial cells have the fern and seed-plant types of PD, respectively. The two SAM organizations may have evolved separately in lycophytes and euphyllophytes and may be associated with gain or loss of the ability to form secondary PD.     

Plastid genome sequencing,comparative genomics,and phylogenomics: Current status and prospects

Abstract More than 190 plastid genomes have been completely sequenced during the past two decades due to advances in DNA sequencing technologies. Based on this unprecedented abundance of data, extensive genomic changes have been revealed in the plastid genomes. Inversion is the most common mechanism that leads to gene order changes. Several inversion events have been recognized as informative phylogenetic markers, such as a 30‐kb inversion found in all living vascular plants minus lycopsids and two short inversions putatively shared by all ferns. Gene loss is a common event throughout plastid genome evolution. Many genes were independently lost or transferred to the nuclear genome in multiple plant lineages. The trnR‐CCG gene was lost in some clades of lycophytes, ferns, and seed plants, and all the ndh genes were absent in parasitic plants, gnetophytes, Pinaceae, and the Taiwan moth orchid. Certain parasitic plants have, in particular, lost plastid genes related to photosynthesis because of the relaxation of functional constraint. The dramatic growth of plastid genome sequences has also promoted the use of whole plastid sequences and genomic features to solve phylogenetic problems. Chloroplast phylogenomics has provided additional evidence for deep‐level phylogenetic relationships as well as increased phylogenetic resolutions at low taxonomic levels. However, chloroplast phylogenomics is still in its infant stage and rigorous analysis methodology has yet to be developed.     

Arbuscular mycorrhizal and septate endophyte fungal associations in lycophytes and ferns of south India

We studied extent and type of arbuscular mycorrhizal (AM) and septate endophytic (SE) fungal associations in five lycophytes and 50 ferns collected from Eastern and Western Ghats regions. Of the 54 species and one variety (belonging to 31 genera) examined; 54 taxa had AM association and AM fungal structures were absent in Marsilea quadrifolia. This is the first report of AM and SE fungal status for 26 species each. Of the 55 taxa examined, AM morphology has been evaluated for the first time in 51 species. The hydrophytic fern Salvinia molesta was mycorrhizal and non-mycorrhizal at different sites. All the epiphytic and saxicolous species examined were mycorrhizal. The percentage of AM colonization ranged from 22.23 (Christella parasitica) to 82.20 (Adiantum lunulatum) in ferns and 53.46 (Selaginella bryopteris) to 84.34 (Selaginella sp.) in lycophytes. Epiphytic life-forms had the maximum average AM colonization levels, whereas aquatic life-forms had the minimum colonization levels. The percentage root length colonized by septate fungi ranged between 0.59 in Ophioglossum reticulatum and 16.36 in Pteris pellucida. The root length with AM and SE fungal structures as well as their total colonization significantly varied among the taxa examined. Most of the lycophytes and ferns had intermediate-type of AM morphology with a few exhibiting Paris-type. AM fungal spore numbers ranged from 1.0 (Angiopteris evecta, Pteridium aquilinum) to (Nephrolepis exaltata) 9.3 spores per 25 g soil and varied significantly among taxa. AM fungal spore morphotypes belonging to Claroideoglomus, Funneliformis, Glomus and Rhizophagus were recorded.     

Diversity of ferns and lycophytes from Reserve Natural Punta Lara, Buenos Aires, Argentina

Punta Lara Natural Reserve shelters the relict of the Southernmost riparian forest in the world, where the flora is threatened by the anthropic expansion and by the introduction of exotic species. In this area the ferns and lycophytes grow in particular environments as marshland, grassland and marginal forest. The aim of this work was to evaluate the diversity of ferns and lycophytes that inhabit in Punta Lara Natural Reserve. Field trips were conducted from 2006 to 2009 and local herbaria and specific bibliography were also revised. As a result of our study twenty four taxa of ferns and lycophytes were recognized. Their habit was mainly terrestrial (70%), fewer aquatic (17%) and epiphytic (13%). Five native taxa were scarcely recorded whereas two native species previously documented were not found. We assume that the main threats on ferns and lycophytes populations in Punta Lara Reserve are: removal for comercialization, invasion by exotic species and climatic fluctuations (drought periods). Keys for taxa identification are given and strategies for the conservation of threatened taxa are proposed.     

Phylogenetic Ordinal Placement Based on rDNA Sequences of the Freshwater Genera Ophioceras and Pseudohalonectria

Chen, W., Shearer, C. A., and Klopp, J. 1995. Phylogenetic ordinal placement based on rDNA sequences of the freshwater genera Ophioceras and Pseudohalonectria. Experimental Mycology 19, 191-201, The ordinal placement of two closely related freshwater genera, Ophioceras and Pseudohalonectria , was assessed by using phylogenetic analysis of morphological characters, partial sequences of the large subunit ribosomal DNA and restriction site variations in the internal transcribed spacer (ITS). The two genera have some morphological features that are used to define taxa in both the Sordariales and Diaporthales, and, hence, their phylogenetic relationships are unclear. Equally weighted analyses of thirty-eight morphological characters produced unresolved phylogenetic trees and unequivocal conclusions could not be drawn based on the morphological data. The polymcrase chain reaction-amplified ITS region was variable in length between the two genera and restriction sites in the ITS region were determined. Analysis of variation in restriction sites in the ITS region placed Ophioceras and Pseudohalonectria in one clade with taxa sampled from Sordariales. About 350 basepairs of DNA sequence from the 5′ end of the large subunit rDNA were also determined. In phylogenetic analysis of the sequence data with Hypocrea lutea and Nectria cinnabarina as outgroups, Ophioceras and Pseudohalonectria showed a closer relationship to Neurospora crassa , Schizothecium sp., and Sordaria fimicola of the Sordariales than to Cryphonectria parasitica and Endothia gyrosa of the Diaporthales.     

Phylogenetic relationships and genetic diversity of the Salicornieae (Chenopodiaceae) native to the Atlantic coasts of France

Phylogenetic relationships of members of the tribe Salicornieae, native to the Atlantic coasts of France, were assessed by three molecular markers: the nuclear ribosomal internal transcribed spacer (ITS), the chloroplast trnL-F and the chloroplast matK sequences. In parallel to the phylogenetic studies, a population genetic study was carried out based on randomly amplified polymorphic DNAs (RAPD). Neither the MP/ML analyses of the sequences, nor the AMOVA and NJ analyses of RAPD fingerprints confirmed the morphology-based classification at the specific level within Salicornia. Instead, our investigations are in favour of the species aggregate concept. Two sister groups were revealed in the genus: one is composed of the diploid taxa, while the other clusters the tetraploid taxa. Conflicting nuclear versus plastid phylogenetic positions of some tetraploid samples, referred to as S. fragilis, indicate that they most likely derive from a reticulate evolution.     

Application of the ITS2 Region for Barcoding Medicinal Plants of Selaginellaceae in Pteridophyta

Background

Selaginellaceae is a family of nonseed plants with special evolutionary significance. Plants of the family Selaginellaceae are similarly shaped and easily confused, complicating identification via traditional methods. This study explored, for the first time, the use of the DNA barcode ITS2 to identify medicinal plants of the Selaginellaceae family.

Methodology/Principal Findings

In our study, 103 samples were collected from the main distribution areas in China; these samples represented 34 species and contained almost all of the medicinal plants of Selaginellaceae. The ITS2 region of the genome was amplified from these samples and sequenced using universal primers and reaction conditions. The success rates of the PCR amplification and sequencing were 100%. There was significant divergence between the interspecific and intraspecific genetic distances of the ITS2 regions, while the presence of a barcoding gap was obvious. Using the BLAST1 and nearest distance methods, our results proved that the ITS2 regions could successfully identify the species of all Selaginellaceae samples examined. In addition, the secondary structures of ITS2 in the helical regions displayed clear differences in stem loop number, size, position, and screw angle among the medicinal plants of Selaginellaceae. Furthermore, cluster analysis using the ITS2 barcode supported the relationship between the species of Selaginellaceae established by traditional morphological methods.

Conclusion

The ITS2 barcode can effectively identify medicinal plants of Selaginellaceae. The results provide a scientific basis for the precise identification of plants of the family Selaginellaceae and the reasonable development of these resources. This study may broaden the application of DNA barcoding in the medicinal plant field and benefit phylogenetic investigations.     

中国现代石松类和蕨类的系统发育与分类系统

石松类和蕨类植物是以孢子繁殖的维管植物,在陆地植物演化上占据重要地位。随着分子系统学研究的开展,各大类群间的系统发育关系得以阐明,传统上的概念得以修正,新的现代石松类和蕨类植物的分类系统也被提出,并不断得到完善。该文介绍国内外在蕨类植物系统发育方面的研究成果,重点讨论中国分布的类群的分类处理。文中提出了一个完整的中国现代石松类和蕨类植物的分类系统,包括5亚纲、14目、39科及12亚科、约140余属。     

Molecular phylogeny and evolution of Caricaceae based on rDNA internal transcribed spacers and chloroplast sequence data

This study focused on clarifying phylogenetic relationships and evolution within Caricaceae. Our phylogenetic analysis based on nucleotide sequences from the ITS of the ribosomal DNA and three chloroplast fragments (matK, trnL-trnF, and psbA-trnH) included 29 taxa belonging to five genera: the neotropical genera Carica, Vasconcellea, Jarilla, and Jacaratia and the equatorial African genus Cylicomorpha. Having a relatively low mutation rate, matK, and trnL-trnF were used for estimating relationships at the generic level, while intrageneric evolution within Vasconcellea was studied with the more variable ITS and psbA-trnH sequences. Gaps, coded as binary characters, were added to the sequence alignments before performing Maximum Parsimony and Maximum Likelihood analyses. Monophyly of Caricaceae as well as phylogenetic distance between Carica and Vasconcellea species, previously belonging to the same genus, and monophyly of the resurrected genus Vasconcellea were emphasized. Within Vasconcellea, the largest genus of this family, two well-confirmed evolutionary lineages could be discerned: (1) V.xheilbornii, V. weberbaueri, V. stipulata, and V. parviflora and (2) a clade holding all other taxa of the genus. Incongruence between nuclear ITS and chloroplast psbA-trnH datasets, shown to be significantly caused by some taxa of the genus Vasconcellea, indicated that reticulate events in this genus might be more frequent than previously suspected. Moreover, intra-individual ITS sequence heterogeneity provided further evidence for the hybrid or introgressed origin of different taxa and one presumed hybrid belonging to this genus.     

Ecological and phylogenetic approaches for diversification of apogamous ferns in Japan

It has long been argued that related asexual and sexual taxa have different distribution patterns. In general, apomictic angiosperms are believed to occur preferentially at higher latitudes and elevations compared to their sexual relatives. It is thus expected that the frequency of apomixis increases with latitude or from warmer to colder climatic regions. However, despite the significant role played by apogamy in fern evolution and diversification, the distribution pattern of apogamous ferns and lycophytes has received little attention. To clarify the ecological diversity pattern and evolutional diversification of apogamous ferns, we analysed a variety of apogamous fern species with reference to latitude, elevation and climatic factors, and reconstructed the ancestral state and estimated the divergence time of Japanese apogamous ferns. Our results on the distribution of apogamous ferns along these two gradients suggest a decline in the proportion of apogamous ferns towards high latitudes and elevations. Temperature was correlated with the proportion of apogamous ferns along both gradients, and the seasonality of precipitation was correlated with the proportion of apogamous ferns along latitude. Reconstruction of ancestral state and estimates of divergence time showed that the crown groups of apogamous ferns diversified less than 15 Ma. The results of our ecological and phylogenetic approaches reinforce the hypothesis based on previously reported phylogenetic results in which the apogamous ferns appears to be correlated with strongly seasonal climates such as the Asia monsoon.