The Indo-Burma biodiversity hotspot for ferns:Updated phylogeny,hidden diversity,and biogeography of the java fern genus Leptochilus(Polypodiaceae)

The Indo-Burma Biodiversity Hotspot is renowned for its rich biodiversity,including that of vascular plants.However,the fern diversity and its endemism in this hotspot have not been well understood and so far,the diversity of very few groups of ferns in this region has been explored using combined molecular and morphological approaches.Here,we updated the plastid phylogeny of the Java fern genus Leptochilus with 226(115%increase of the latest sampling)samples across the distribution range,specifically those of three phylogenetically significant species,Leptochilus ovatus,L.pedunculatus,and L.pothifolius.We also reconstructed the first nuclear phylogeny of the genus based on pgiC gene data.Based on molecular and morphological evidence,we identified three new major clades and six new subclades,redefined three existing species,discovered a number of cryptic species of the genus,and elucidated the evolution of the three most variable characters.Our divergence time analyses and ancestral area reconstruction showed that Leptochilus originated in the Oligocene and diversified from early Miocene and 15 dispersal events from lower to higher latitudes are identified.The evolution of three most important morphological characters is analyzed in a context of the new phylogeny.Our analysis showed that 30(59%of total 51)species of Leptochilus occur in Indo-Burma hotspot,24(80%of the 30 species)of which are endemic to this hotspot.We argue that the Indo-Burma hotspot should be recognized as a diversity hotspot for ferns.

Resources of Wild Ornamental Ferns in Guizhou Province and Their Utilization

Through investigating the wild ornamental ferns in Guizhou Province, it is found that there are totally 312 species (including variations), 111 genera and 49 families of such resources in this region, which are classified into terrestrial fern, epiphytic fern, saxicolous fern, vine fern, tree fern and aquatic fern according to their ecological types. The application of ornamental ferns in garden landscaping and development countermeasures are analyzed in this study.

Resource Distribution and Exploitation of Edible Ferns in Guizhou

64 species （including 5 varieties）, 30 genera and 24 families of the edible ferns in Guizhou are preliminartiy confirmed, according to the combination of spot investigation and refereuce analySis：the species, genera and families account for 8.13%, 19.74% and 44.44% of ferns in Guizhou, respectively. The floristic composition characters and resource distribution of 23 kinds of the edible ferns in Guizhou are described.

Prediction of Suitable Habitat for Lycophytes and Ferns in Northeast China: A Case Study on Athyrium brevifrons

Suitable habitat is vital for the survival and restoration of a species.Understanding the suitable habitat range for lycophytes and ferns is prerequisite for effective species resource conservation and recovery efforts.In this study, we took Athyrium brevifrons as an example, predicted its suitable habitat using a Maxent model with 67 occurrence data and nine environmental variables in Northeast China.The area under the curve(AUC) value of independent test data, as well as the comparison with specimen county areal distribution of A.brevifrons exhibited excellent predictive performance.The type of environmental variables showed that precipitation contributed the most to the distribution prediction, followed by temperature and topography.Percentage contribution and permutation importance both indicated that precipitation of driest quarter(Bio17) was the key factor in determining the natural distribution of A.brevifrons, the reason could be proved by the fern gametophyte biology.The analysis of high habitat suitability areas also showed the habitat preference of A.brevifrons: comparatively more precipitation and less fluctuation in the driest quarter.Changbai Mountains, covering almost all the high and medium habitat suitability areas, provide the best ecological conditions for the survival of A.brevifrons, and should be considered as priority areas for protection and restoration of the wild resource.The potential habitat suitability distribution map could provide a reference for the sustainable development and utilisation of A.brevifrons resource, and Maxent modelling could be valuable for conservation management planning for lycophytes and ferns in Northeast China.

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 2 C 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 2 C 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.

Global patterns of fern species diversity:An evaluation of fern data in GBIF

Despite that several studies have shown that data derived from species lists generated from distribution occurrence records in the Global Biodiversity Information Facility(GBIF)are not appropriate for those ecological and biogeographic studies that require high sampling completeness,because species lists derived from GBIF are generally very incomplete,Suissa et al.(2021)generated fern species lists based on data with GBIF for 100 km×100 km grid cells across the world,and used the data to determine fern diversity hotspots and species richness-climate relationships.We conduct an evaluation on the completeness of fern species lists derived from GBIF at the grid-cell scale and at a larger spatial scale,and determine whether fern data derived from GBIF are appropriate for studies on the relations of species composition and richness with climatic variables.We show that species sampling completeness of GBIF is low(<40%)for most of the grid cells examined,and such low sampling completeness can substantially bias the investigation of geographic and ecological patterns of species diversity and the identification of diversity hotspots.We conclude that fern species lists derived from GBIF are generally very incomplete across a wide range of spatial scales,and are not appropriate for studies that require data derived from species lists in high completeness.We present a map showing global patterns of fern species diversity based on complete or nearly complete regional fern species lists.

Diversity, Endemism and Conservation of Ferns(Polypodiales) in the Mexican Mountain Component

We analyzed the distributional patterns of95 selected species of leptosporangiate ferns inhabiting the Mexican Mountain Component, using grid-cells of one geographical degree as unit areas,applying endemism indices and richness and beta diversity analyses.Distributional data were obtained from several herbaria and specialized literature.Five grid-cells appear to be important for fern species richness, as they contain 35 to 49 species.These gridcells are located in the Sierra Madre Oriental(SMO),Trans-Mexican Volcanic Belt(TMVB) and the Sierra Madre del Sur(SMS).Mean richness by latitudinal belts of one degree showed that the belts with highest values are related to the TMVB and SMS.A total of 13grid-cells were recognized as important from theperspective of endemism; most of them are located also in the SMO, TMVB and SMS.The richest gridcells coincided with one of the main centres of endemism for ferns obtained in this study, located in the convergence of the southern part of the SMO, the eastern portion of the TMVB and the northern part of the SMS, reflecting the high humidity existing on the mountain slopes facing the Gulf of Mexico.Some important grid-cells recognized from richness and endemism analyses coincide with Mexican Natural Protected Areas.The beta diversity analysis showed a low degree of similarity among grid-cells, implying a high species replacement, as the result of environmental heterogeneity occurring in the Mexican mountain systems.On the other hand, the spatial analysis suggested a pattern of phytogeographical regionalization comprising twomain areas: the Mexican Transition Zone and the Mexican Central Plateau.Ferns play an important role in the Mexican biodiversity and contributing to the beta diversity of Mexico.

Fern diversity and biomass at Chilapatta reserve forest of West Bengal Terai Duars in sub-humid tropical foothills of Indian eastern Himalayas

We documented the status of fern diversity, biomass and car- bon accumulation at Chilapatta Reserve Forest in Cooch Behar Wildlife Division, West Bengal. Stratified random nested quadrat sampling was adopted for analyzing the qualitative and quantitative characters. Nineteen fern species were recorded, of which two are yet to be identified. Identified species were of eight families and nine genera. Highest and lowest frequency recorded were 25.44 and 0.19 while relative frequency varied from 3.16 to 12.25. Fern density ranged from 93 to 13,403 individuals.ha^-1. Most of the species were widely distributed. IVI values ranged from 7.54 to 37.45. The above ground portion of ferns accumulated the major portion ofbiomass and carbon.

Fern extracts potentiate fluconazole activity and inhibit morphological changes in Candida species

Objective: To investigate the antifungal activity of the fern species Lygodium venustum(L. venustum) and Pityrogramma calomelanos(P. calomelanos) against Candida albicans and Candida tropicalis strains.Methods: The microdilution method was used to evaluate the antifungal activity, as well as the modulating effects of ethanolic extracts of these plants in combination with fluconazole. The minimum inhibitory concentration(MIC), minimum fungicide concentration and morphological changes were also determined.Results: The extract obtained from L. venustum presented a MIC > 8 192 mg/m L, while the extract obtained from and P. calomelanos presented a MIC = 8 192 mg/m L, indicating that they present weak antifungal activity. However, combination of the extracts with Fluconazole potentiated the antifungal activity of this drug. At different experimental conditions, such as concentration of the extract and type of strain, the extracts inhibited hyphae and pseudohyphae formation, indicating that these fern species can affect the morphology of the fungi.Conclusions: The extracts obtained from the fern species L. venustum and P. calomelanos dose not present significant antifungal activity. However, P. calomelanos potentiates the activity of fluconazole and both extracts inhibits the morphological changes in Candida species, indicating that they have potential pharmacological activity as modulators of fungal biology. Therefore, novel studies are required to characterize the interference of these extracts in the virulence and pathogenicity of Candida species as well as the potential of fern species to treat fungal infections.

Fern Flora in Grassland of Gansu Province

In order to fully understand the distribution characteristics and geographical elements of fern flora and to reveal their process of natural history, the present study analyzed the floristic composition and geographical distribution of ferns in grassland of Gansu Province. The results showed that there were 31 species of ferns in grassland of Gansu, belonging to 19 genera and 14 families. The investigated ferns accounted for 22.22%, 8.26% and 1.99% of total families （63）, total genera （230） and total species （2 600） of ferns resources, respectively. The dominant families were Dryopteridceae and Polypodiaceae; the dominant genera were Selaginella and Woodsia. Families mainly belonged to tropical distribution type and genera mainly belonged to tropical and temperate distribution type without endemic genus. Vertical distribution was obvious ; plant components contained old families and genera. The richness of fern in Gansu grassland was not high.

High Frequency Sporophytes Regeneration from the Spore Culture of the Endangered Aquatic Fern <i>Isoetes coreana</i>

Using a mixed culture of megaspores and microspores from I. coreana, we established high frequency sporophyte regeneration system. After 20 days of culturing in MS basal medium, microscopic examination showed significant morphological changes and the microspore released numerous small vesicles into the culture medium. Megaspores also showed dramatic morphological changes during its incubation time in culture. The spore wall was cracked by the expansion of the megaspore (about 2 times increase in diameter). Simultaneously, brown spots were observed on the surface of the megaspores. The frequency of female gametophytes developing from immature megaspores cultured in MS basal liquid medium (pH 7) supplemented with 1 mgl-1 GA3 was 46%. However, these female gametophytes derived from megaspore only culture could not differentiate into sporophytes. The mixed culture of microspores and megaspores resulted in successful sporophyte regeneration. The highest frequency (12.3%) of green sporophyte regeneration from mixed spore culture occurred when the cultures were maintained at 25℃ under cool-white fluorescent light (40 μmol·m-2·s-1) with a 16 h photoperiod. Regenerated sporophytes were transferred to a test tube containing vermiculite and a sand mixture and left there until they had three leaves. After root growth and the fifth leaf had emerged, more than 95% of the regenerated sporophytes were successfully transferred to the soil and grown to mature plants. The sporophyte regeneration system established in this study could be successfully used for the restoration of the endangered aquatic species, I. coreana.

A computer-based image analysis for tear ferning featuring

The present work focuses on the development of a novel computer-based approach for tear ferning(TF)featuring.The original TF images of the recently developedfive-point grading scale have been used to assign a grade for any TF image automatically.A vector characteristic(VC)representing each grade was built using the reference images.A weighted combination between features selected from textures analysis using gray level co-occurrence matrix(GLCM),power spectrum(PS)analysis and linear specificity of the image were used to build the VC of each grade.A total of 14 features from texture analysis were used.PS at di®erent frequency points and number of line segments in each image were also used.Five features from GLCM have shown significant di®erences between the recently developed grading scale images which are:angular second moment at 0
and 45
,contrast,and correlation at 0
and 45
;thesefive features were all included in the characteristic vector.Three specific power frequencies were used in the VC because of the discrimination power.Number of line segments was also chosen because of dissimilarities between images.A VC for each grade of TF reference images was constructed and was found to be significantly different from each other's.This is a basic and fundamental step toward an automatic grading for computer-based diagnosis for dry eye.

Optimization of Conditions for Extracting Luteolin from Ternate Grape Fern Herb by an Orthogonal Experiment

[Objectives] This study was conducted to optimize the extraction process of luteolin, a natural product of Ternate Grape Fern Herb, so as to provide a theoretical research basis for the chemical composition, pharmacological research and preparation development of Ternate Grape Fern Herb. [Methods] The effects of four factors including ultrasonic extraction time, liquid-to-material ratio, ultrasonic extraction temperature and ethanol concentration on extraction yield of luteolin from Ternate Grape Fern Herb were investigated by the ultrasonic-assisted extraction technology, through a single-factor experiment and an orthogonal experiment based on the single-factor experiment, and the experimental data was subjected to variance analysis and SSR analysis of the significance of differences between groups by SPSS20.0 software to select the optimal extraction process conditions. [Results] The optimal extraction process conditions were as follows: ultrasonic extraction time 25 min, liquid-to-material ratio 25∶1 ml/g, ethanol volume fraction 60%, ultrasonic extraction temperature 85 ℃. [Conclusions] The results were verified, and it was determined that the ultrasonic-assisted extraction process established in this study is reasonable and reliable, and can be used for the extraction of luteolin.

CLINICAL APPLICATION AND EVALUATION OF SALIVA FERNING TEST IN SJGREN' S SYNDROME

Objective To evaluate the saliva ferning test (SFT) as diagnostic test for xerostomia in patients with Sjogren’s Syndrome (SS). Methods In this study, dried samples of freshly produced saliva from 78 patients with established SS according to European Community criteria and 80 healthy controls were examined by light microscopy. The crystallization was classified into 4 types according to the ferning phenomenon: uniformity, branching, spreading and integrity (type I normal and type Ⅱ,Ⅲ,Ⅳ abnormal ). Then, the 78 patients underwent lebial salivary gland biopsy. According to Tarpley’s classifica- tion, minor salivary gland biopsy (≥2 + ) was considered to be positive . Results 1. The sensitivity of SFT was high (70/ 78 = 89. 74% ). And the specificity wes also high (67/80 = 83. 75% ). 2. Abnormal SFT was observed in 70/78 (89. 74% ) samples from patients group and in 13 /80 (16. 25% ) samples from healthy controls. The differences of SFT in patients group versus controls were statistically significant (P< 0. 01 ). 3. The sensitivity of SFT and mini labial gland biopsy had no significant differences (P > 0. 05) as diagnostic tests in SS. Conclusion SFT was simple, sensitive and specific as diagnostic test in SS suspect patients just as mini labial gland biopsy.

Elaphoglossum(Dryopteridaceae-Fern)of Amazon Rainforest in Brazil:Anatomic Characterization and Adaptative Strategies

This study describes the anatomy of sterile leaves of Elaphoglossum discolor (Kuhn) C. Chr., E. flaccidum (Fée) T. Moore and E. laminarioides (Bory ex Fée) T. Moore, the most representative species of the genus in the Ecological Park of Gunma in Pará State. It reports the main diagnostic characters and provides new systematic data for the group. In addition, it locates the production and accumulation sites of bioactive compounds to determine possible adaptive strategies of these species in the Amazon rainforest environment. Diagnostic structural features include stoma typology, central veins and margin forms, type of mesophyll, and the presence of schlerenchymatous sheaths in the cortex, among others. Among the bioactive compounds related to defense adaptation are phenolic compounds, which occur in all three species, and alkaloids and mucilage, which are exclusive to E. laminarioides. Of the three species studied, E. laminarioides has features that make it the best suited to the rainforest environment.

福建汀江源自然保护区石松类和蕨类植物区系分析

【目的】探讨汀江源自然保护区石松类和蕨类植物从物种多样性和区系地理特征,以期为两类植物多样性保护、资源应用开发及系统演化等研究提供参考。【方法】在野外调查、标本鉴定基础上,结合文献资料,整理该区石松类和蕨类植物名录,分析区系组成及地理成分。【结果】(1)该区石松类和蕨类植物有23科60属129种,其中石松类有2科5属15种,蕨类植物有21科55属114种。(2)优势科、属明显,优势科有3科,优势属有1属;单种科、属和寡种科、属是构成该区石松类和蕨类植物科、属的主体。(3)科的分布区类型以世界分布和热带分布为主;属、种的分布区类型以热带分布为主,同时呈现温带性质;中国特有种有15种。(4)该区与梁野山保护区的地理亲缘关系最近。【结论】汀江源国家级自然保护区石松类和蕨类植物地理成分多样,特有现象明显,生物多样性较低,植物区系成分在水平梯度上呈现由热带向温带过渡的特征。

广东车八岭国家级自然保护区石松类和蕨类植物多样性研究

为全面了解广东车八岭国家级自然保护区的植物资源,并为区内生物多样性的长期监测、保护管理和科普教育提供数据支持,于2023年6月至8月通过样线法对该区石松类和蕨类植物进行野外考察和标本采集,并结合文献资料对保护区内石松类和蕨类植物的物种组成、区系特征及资源情况进行统计分析。结果表明:(1)车八岭保护区共有石松类和蕨类植物25科75属231种(含种下等级),本次调查新增保护区内植物64种,隶属于11科28属。(2)保护区内石松类和蕨类植物有单种科6科、单种属38属,分别占总科、属数的24.00%和50.67%,表明该区石松类和蕨类植物具有一定的古老性和孑遗性。(3)保护区内石松类和蕨类植物的区系地理成分复杂多样,其中科属以热带成分为主,分别占总科、属数的60.00%和68.00%;物种以亚热带成分为主(54.98%),热带成分次之(29.87%),符合亚热带植物区系的特征,并具有一定程度的热带区系性质。(4)与周边保护区相比较,车八岭保护区与青云山省级自然保护区的关系最为密切,科属种的相似性指数(Jaccard/Krober系数)分别为92.00/96.00、73.42/85.33、42.42/62.86,推测与两者地理距离较近和气候特征相似有关。(5)保护区内石松类和蕨类植物资源丰富,其中具食用价值6种、药用价值145种、观赏价值141种和工业价值32种,并发现4种国家重点保护石松类和蕨类植物。

观赏蕨类姬蕨同配子体自交的孢子体萌发和发育特性

[目的]评估交配形式对蕨类繁殖的影响。[方法]以姬蕨(Hypolepis punctata)为例,设计配子体单独培养(同配自交)和混合培养(混交)比较试验,比较两种交配系统下孢子体萌发状态(萌发率、存活率),孢子体成长情况及生长速率。[结果]尽管自交幼苗生长趋势弱于混交个体,单独培养产生的完全纯合的姬蕨孢子体最终存活率达到62.5%,说明蕨类的自交确实可以导致近交衰退,但仍然具有可接受的存活能力,单个孢子仍有很大概率上延续种群。蕨类植物由于其特殊的生活史特性,可通过单独培养得到完全纯合的自交个体,从而得到性状稳定的品系。[结论]该研究中单独培养技术在园艺育种方向具有很好的应用前景。