Chromosome study of the fern genus Cyrtomium (Dryopteridaceae)

Cyrtomium (Dryopteridaceae) is an Asiatic genus comprising about 40 species characterized by having anastomosing veins that form areolae with included veinlets. Twelve chromosome counts representing 11 taxa were made in the present study. The chromosome numbers of six species endemic to China are reported for the first time: C. uniseriale , 2 n  = 164; C. guizhouense , 2 n  = 82; C. shingianum , n  = 82, 2 n  =  c .164; C. chingianum , 2 n  = 164; C. urophyllum , 2 n  = 82 and C. aequibasis , 2 n  = 123. The results were used to show that (1) C. uniseriale should be reduced to varietal status under C. balansae ; (2) subseries Balansana is not closely related to any other Cyrtomium species and should be separated from Cyrtomium ; and (3) C. guizhouense and C. lonchitoides are basal groups when subseries Balansana is not considered. These results were also supported by morphology. A high level of differentiation is a distinct characteristic in Cyrtomium . The genus has two reproductive types: sexual and apogamous. Sexual species are distributed mostly in China. Southwestern China is a modern diversity centre of Cyrtomium , with 80.6% of known species occurring there. Many species are endemic, most of which are sexual tetraploids or apogamous triploids. Both the apogamous reproductive type and polyploid species are adaptions to limestone habitats.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 150 , 221–228.     

Direct production of sporophytic plants from spores of Equisetum arvense

Spores of Equisetum arvense were cultured in Murashige and Skoog liquid medium with or without cytokinin. Without cytokinin, spores germinated 2–3 days after sowing and developed into normal young gametophytes. These gametophytes were composed of well-vacuolated cells. On the other hand, germinated spores in the medium with cytokinin formed globular cell masses that were composed of small and dense cells. These cell masses developed into sporophytic plants after further culture. This revised version was published online in June 2006 with corrections to the Cover Date.     

傅氏凤尾蕨配子体发育及其无配子生殖的研究

采用光学显微镜对傅氏凤尾蕨（Pteris fauriei Hieron.）的配子体发育进行了研究。傅氏凤尾蕨孢子深褐色,呈四面体形,三裂缝,极面观为钝三角形,赤道面观近半圆形。孢子自播种3～7 d左右开始萌发,萌发类型为书带蕨型。18 d左右发育为丝状体,大约为6～10个细胞长;25 d左右,丝状体可通过顶端细胞或中间细胞分裂产生片状体。播种40～50 d左右,在片状体的侧面产生分生组织,发育成原叶体,发育类型为水蕨型。雄配子体形状不规则,其上可产生精子器。大型原叶体形状较规则,顶端为心形,发育2～3个月左右,在生长点下方产生无性胚芽,进行无配子生殖。此后不久,在胚芽和生长点之间产生导管连接孢子体和配子体。经多次培养表明傅氏凤尾蕨不产生颈卵器,为专性无配子生殖。     

A current perspective on apomixis in ferns

This review provides a synopsis of apogamous reproduction in ferns and highlights important progress made in recent studies of fern apomixis. First, a summary of the apomictic fern life cycle is provided, distinguishing between two pathways to diploid spore production that have been documented in apomictic ferns (premeiotic endomitosis and meiotic first division restitution) and briefly discussing the evolutionary implications of each. Next, recent trends in fern apomixis research are discussed, exposing a shift in focus from the observation and characterization of apomixis in ferns to more integrated studies of the evolutionary and ecological implications of this reproductive mode. Peer-reviewed contributions from the past decade are then summarized, spanning the identification of new apomictic lineages through to the developmental, phylogenetic, and population genetic insights that have been made in studies of fern apomixis during that time. Gaps in our understanding are also discussed, including the extent and implications of recombinant apomixis in ferns, the possible reversibility of reproductive mode (from apomictic to sexual) in ferns, and the genomic causes and consequences of apomixis in seed free vascular plants. To conclude, future directions for fern apomixis research are proposed in the context of modern technological advances and recent insights from studies of apomixis in other groups.     

Spore germination and isozyme patterns in the apomictic fern Cyrtotnium falcatum

Progeny from a plant of the apomictic species Cyrtomium falcatum were examined by protein electrophoresis. These progeny were produced from spores originating in sporangia with differing spore numbers. Analysis of zymograms indicates that variant banding patterns were exhibited in 16.1% of the progeny sampled. These anomalous phenotypes suggest that some apomictically reproducing plants can produce genetically variable progeny.     

AN EVALUATION OF METHODS USED TO ASSESS INTERGENERIC HYBRIDIZATION IN KELP USING PACIFIC LAMINARIALES (PHAEOPHYCEAE)1

Kelp intergeneric laminarialean hybridizations and hybridization protocol were assessed using seven northeast Pacific kelp species: Alaria marginata Postels and Ruprecht, Costaria costata (C. A. Agardh) Saunders, Eisenia arborea Areschoug, Laminaria saccharina (L) Lamouroux, Lessoniopsis littoralis (Tilden) Reinke, Macrocystis integrifolia Bory, and Nereocystis leutkeana (Mertens) Postels and Ruprecht. Survival and development of sporophyte morphologies derived from selfings, separate males and females, and reciprocal crosses were evaluated over 30 weeks of cultivation. All cultures were initiated from cloned gametophytes. Two closely related species, Laminaria angustata Kjellman and L. japonica Areschoug, demonstrated the efficacy of long‐term (up to 30 years) cloned gametophytes in hybridization studies. Sporophyte morphologies appeared in 34%–69% of control and hybridization trials, and 6%–16% of all trials produced sporophytes in control and hybridization conditions that persisted through 30 weeks of cultivation. Sporophytes in control and hybridization conditions could appear normal or abnormal. Usually, the morphology of sporophytes in hybridizations and female controls resembled the female parent, whereas the sporophytes in male controls often had an abbreviated morphology, lacking definitive generic features. Species‐specific rDNA internal transcribed spacer molecular primers were used to determine the parentage of five putative hybrids. Only the L. japonica♀/L. angustata♂ hybrid bore both parental genomes. That negative controls could produce persistent and normal‐appearing sporophytes negates their value and emphasizes the importance of molecular confirmation in hybridization studies. These findings were applied to critique the only known wild intergeneric hybrid, Pelagophycus/Macrocystis.     

鳞毛蕨科四个属的染色体数目

报道了鳞毛蕨科的4个属(毛枝蕨属Leptorumohra、石盖蕨属Lithostegia、黔蕨属Phanerophlebiopsis和柳叶蕨属Cyrtogonellum)6种植物的染色体数目及生殖方式。其中柳叶蕨Cyrtogonellum fraxinellum 'n'=123, 2n=123; 离脉柳叶蕨C. caducum 'n'=123; 斜基柳叶蕨C. inaequalis 2n=123; 四回毛枝蕨Leptorumohra quadripinnata n=41; 石盖蕨Lithostegia foeniculacea 2n=164; 长叶黔蕨Phanerophlebiopsis neopodophylla 2n=82。石盖蕨属、黔蕨属和柳叶蕨属的染色体数目为首次报道。结果表明这3个属的染色体基数和鳞毛蕨科中其他属一样均为x=41。细胞学证据支持将石盖蕨属、黔蕨属置于鳞毛蕨科的处理。本文还发现柳叶蕨属与贯众属Cyrtomium的一些种一样, 具有无融合生殖方式, 而其他3个属仅具有在蕨类植物中较为常见的有性生殖方式。     

金粉蕨配子体发育及其无配子生殖的研究

采用光学显微镜和半薄切片技术对金粉蕨(Onychium siliculosum(Desv.)C.Chr.)的配子体发育及其无配子生殖进行了研究。金粉蕨孢子黄褐色,具三裂缝,孢子萌发时先产生假根,然后长出原叶体细胞,萌发类型为书带蕨型。原叶体细胞经丝状体、片状体发育为原叶体,发育类型为水蕨型。金粉蕨配子体分枝发达,成熟配子体呈丛状。多次培养表明金粉蕨只能产生精子器,不能产生颈卵器,为专性无配子生殖。半薄切片观察表明配子体生长点下方的细胞经分裂产生一群小型细胞,由它们发育为无配子生殖胚。小型细胞先分化出无性胚芽,再分化出胚根。胚芽与胚根,胚与配子体之间由分化出的导管连接。     

毒莠定作为外源激素促进水稻子房培养中胚状体的分化

用毒莠定(4-氨基-3,5,6-三氯-2-羧基吡啶)代替MCPA(2-甲基-4-氯苯氧乙酸)作为水稻未传粉子房培养中的外源激素,所诱导的助细胞无配子生殖胚状体分化率大幅度提高,愈伤组织化的倾向大为减弱,并观察到在诱导培养基上直接出苗的现象。用N_6基本培养基附加3%的蔗糖、2毫克/升的毒莠定与500毫克/升的水解乳蛋白可获得较佳的培养效果。组织学观察表明在毒莠定的作用下,原胚多数是沿着接近体内合子胚正常分化的途径形成具根、芽的两极结构,而和MCPA所诱导的原胚不分化与愈伤组织化有很大的差异。讨论了毒莠定作为外源激素在植物组织培养中应用的潜力。     

Premeiotic endomitosis and the costs and benefits of asexual reproduction

Different patterns of development can influence the strength and direction of selection of a trait. Here, it is shown how this may be the case for asexual reproduction in which eggs (animals) or spores (plants) inherit the maternal chromosome number through a precursor cell undergoing an endoduplication of its chromosomes prior to meiotic reduction. Asexuality involving premeiotic endoduplication (APE) has a wide but uneven distribution among animals and plants. It is argued that different patterns of egg and spore production and differences in when endoduplication occurs, together with differences in breeding structure (dioecious vs. hermaphroditic) and reproductive strategy (e.g. spawning vs. vivipary), can result in APE being associated with fecundity costs that can result in an overall cost of sex or in an overall cost of asex or in APE being selectively neutral relative to sex. There is a lack of a close correlation between the taxonomic distribution of APE and its potential costs and benefits however, possible causes of which are explored. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 109 , 487–495.     

Apogamous sporophyte formation in a fernPteris multifida and its characteristics

When spores of the fern,Pteris multifida, were aseptically cultured in the dark, sporophytic plants were apogamously induced. The plants have been subsequently grown in pots until the development of leaves with many sporangia for observations of meiotic characteristics in their sporocytes. The sporophytic plants originated from spores were estimated to be haploid, and the estimation was supported by abnormal meiosis in sporocytes and the absence of mature spores, but some chromosomes (n=58) formed bivalents in the meiotic process.     

Induced apogamous sporophytes inPteris dispar andP. semipinnata,and the meiotic behavior in their sporocytes

Spores ofPteris dispar andP. semipinnata were aseptically cultured in flasks for apogamous sporophyte induction. Calli or cell colonies similar to calli were induced in cultures supplemented with hormones. Sporophytic leaves subsequently developed from them in hormone-free medium and the young sporophytes were raised into plants with sporangia. Since the wild-type plants having 116 chromosomes are tetraploid, the sporophytic plants originating from spores would appear to be diploid (dihaploid). In induced sporophytes ofP. semipinnata, non-homologous chromosomes (58 univalents) were found during the meiotic process in sporocytes. InP. dispar, however, the meiotic cells showed many bivalent chromosomes (maximum 29ll). These results suggest thatP. semipinnata is allotetraploid, whereasP. dispar is autotetraploid.     

Apomixis and reticulate evolution in the Asplenium monanthes fern complex

Background and Aims

Asexual reproduction is a prominent evolutionary process within land plant lineages and especially in ferns. Up to 10 % of the approx. 10 000 fern species are assumed to be obligate asexuals. In the Asplenium monanthes species complex, previous studies identified two triploid, apomictic species. The purpose of this study was to elucidate the phylogenetic relationships in the A. monanthes complex and to investigate the occurrence and evolution of apomixis within this group.

Methods

DNA sequences of three plastid markers and one nuclear single copy gene were used for phylogenetic analyses. Reproductive modes were assessed by examining gametophytic and sporophyte development, while polyploidy was inferred from spore measurements.

Key Results

Asplenium monanthes and A. resiliens are confirmed to be apomictic. Asplenium palmeri, A. hallbergii and specimens that are morphologically similar to A. heterochroum are also found to be apomictic. Apomixis is confined to two main clades of taxa related to A. monanthes and A. resiliens, respectively, and is associated with reticulate evolution. Two apomictic A. monanthes lineages, and two putative diploid sexual progenitor species are identified in the A. monanthes clade.

Conclusions

Multiple origins of apomixis are inferred, in both alloploid and autoploid forms, within the A. resiliens and A. monanthes clades.     

Analysis of nucleolar organizer constitution by fluorescent in situ hybridization (FISH) in diploid and artificially produced haploid sporophytes of the fernOsmunda japonica (Osmundaceae)

HaploidOsmunda japonica (2n = × = 22), produced in tissue culture by induced apogamy and acclimatized in vivo, was cytologically compared with the diploid sister plant. The haploid had smaller guard cells than the diploid and a numerically exactly reduced karyotype. Fluorescent in situ hybridization (FISH) using an rDNA probe showed four hybridization signals (one large and three small ones) in the haploid and eight signals (two large and six small ones) in the diploid plant. These results represent a cytological proof for the origin of the haploid plant by apogamy without recognizable chromosome aberrations.