Diazotrophic community structure and function in two successional stages of biological soil crusts from the Colorado Plateau and Chihuahuan Desert

The objective of this study was to characterize the community structure and activity of N2-fixing microorganisms in mature and poorly developed biological soil crusts from both the Colorado Plateau and Chihuahuan Desert. Nitrogenase activity was approximately 10 and 2.5 times higher in mature crusts than in poorly developed crusts at the Colorado Plateau site and Chihuahuan Desert site, respectively. Analysis of nifH sequences by clone sequencing and the terminal restriction fragment length polymorphism technique indicated that the crust diazotrophic community was 80 to 90% heterocystous cyanobacteria most closely related to Nostoc spp. and that the composition of N2-fixing species did not vary significantly between the poorly developed and mature crusts at either site. In contrast, the abundance of nifH sequences was approximately 7.5 times greater (per microgram of total DNA) in mature crusts than in poorly developed crusts at a given site as measured by quantitative PCR. 16S rRNA gene clone sequencing and microscopic analysis of the cyanobacterial community within both crust types demonstrated a transition from a Microcoleus vaginatus-dominated, poorly developed crust to mature crusts harboring a greater percentage of Nostoc and Scytonema spp. We hypothesize that ecological factors, such as soil instability and water stress, may constrain the growth of N2-fixing microorganisms at our study sites and that the transition to a mature, nitrogen-producing crust initially requires bioengineering of the surface microenvironment by Microcoleus vaginatus.     

Temporal Variation in Community Composition, Pigmentation, and Fv/Fm of Desert Cyanobacterial Soil Crusts

Summers on the Colorado Plateau (USA) are typified by harsh conditions such as high temperatures, brief soil hydration periods, and high UV and visible radiation. We investigated whether community composition, physiological status, and pigmentation might vary in biological soil crusts as a result of such conditions. Representative surface cores were sampled at the ENE, WSW, and top microaspects of 20 individual soil crust pedicels at a single site in Canyonlands National Park, Utah, in spring and fall of 1999. Frequency of cyanobacterial taxa, pigment concentrations, and dark adapted quantum yield [F(v)/F(m)] were measured for each core. The frequency of major cyanobacterial taxa was lower in the fall compared to spring. The less-pigmented cyanobacterium Microcoleus vaginatus showed significant mortality when not in the presence of Nostoc spp. and Scytonema myochrous (Dillw.) Agardh. (both synthesizers of UV radiation-linked pigments) but had little or no mortality when these species were abundant. We hypothesize that the sunscreen pigments produced by Nostoc and Scytonema in the surface of crusts protect other, less-pigmented taxa. When fall and spring samples were compared, overall cyanobacterial frequency was lower in fall, while sunscreen pigment concentrations, chlorophyll a concentration, and F(v)/F(m) were higher in fall. The ratio of cyanobacterial frequency/chlorophyll a concentrations was 2-3 times lower in fall than spring. Because chlorophyll a is commonly used as a surrogate measure of soil cyanobacterial biomass, these results indicate that seasonality needs to be taken into consideration. In the fall sample, most pigments associated with UV radiation protection or repair were at their highest concentrations on pedicel tops and WSW microaspects, and at their lowest concentrations on ENE microaspects. We suggest that differential pigment concentrations between microaspects are induced by varying UV radiation dosage at the soil surface on these different microaspects.     

古尔班通古特沙漠生物结皮不同发育阶段中藻类的变化

通过在古尔班通古特沙漠南缘相同的地貌部位,选择裸沙、藻结皮、地衣结皮和苔藓结皮4种不同演替阶段中的生物结皮,研究了藻类的种类组成、优势种和生物量的变化.结果表明:(1)在结皮的不同演替阶段,藻类种类组成不同,其常见物种有一定的差异,如裸沙中藻类常见种是脆杆藻2(Fragilaria sp.2)、威利颤藻(Oscillatoria willei)和奥克席藻(Phormidium okenii),藻结皮的常见种是小聚球藻(Synechococcus parvus)、颗粒常丝藻 (Tychonema granulatum)、韧氏席藻(Phormidium retzli);同时在不同发育阶段亦存在一些特有种.(2)在裸沙发育到成熟生物结皮的过程中,藻类的优势物种也发生相应的变化.裸沙、藻结皮、地衣结皮和苔藓结皮的优势种分别是脆杆藻1(Fragilaria sp.1)、具鞘微鞘藻(Microcoleus vaginatus)、具鞘微鞘藻、眼点伪枝藻(Scytonema ocellatum)或集球藻(Palmellococcus miniatus).(3)藻类生物量在生物结皮不同演替阶段差异极显著(P<0.01),在裸沙中藻类生物量最低,随着生物结皮的逐渐发育,藻类生物量明显升高,地衣结皮最高,约是裸沙的8.3倍,当发育至苔藓结皮时,藻类生物量又有所下降.(4)在裸沙中基本为松散的沙粒,随着生物结皮的演替,丝状种类占明显的优势,尤其是具鞘微鞘藻,另外真菌菌丝和苔藓假根分别在地衣结皮和苔藓结皮中起着重要作用.     

Molecular characterization of hot spring cyanobacteria and evaluation of their photoprotective compounds

Phylogenetic analysis of 4 cyanobacterial strains isolated from hot springs in Rajgir, India, was carried out using the 16S rRNA gene (1400 bp). These strains were identified as members of Chroococcales ( Cyanothece sp. strain HKAR-1) and Nostocales ( Nostoc sp. strain HKAR-2, Scytonema sp. strain HKAR-3, and Rivularia sp. strain HKAR-4). Furthermore, we evaluated the presence of ultraviolet-screening and (or) photoprotective compounds, such as mycosporine-like amino acids (MAAs) and scytonemin, in these cyanobacteria by using high-performance liquid chromatography. Well-characterized MAAs, including the critical and highly polar compounds shinorine, porphyra-334, and mycosporine-glycine, as well as several unknown MAAs, were found in these hot-spring-inhabiting microorganisms. The presence of scytonemin was detected only in Scytonema sp. strain HKAR-3 and Rivularia sp. strain HKAR-4. The results indicate that hot spring cyanobacteria, namely Cyanothece, Nostoc, Scytonema, and Rivularia, belonging to different groups possess various photoprotective compounds to cope up with the negative impacts of damaging radiations.     

Do lichens domesticate photobionts like farmers domesticate crops? Evidence from a previously unrecognized lineage of filamentous cyanobacteria

Phylogenetic diversity of lichen photobionts is low compared to that of fungal counterparts. Most lichen fungi are thought to be associated with just four photobiont genera, among them the cyanobacteria Nostoc and Scytonema, two of the most important nitrogen fixers in humid ecosystems. Although many Nostoc photobionts have been identified using isolated cultures and sequences, the identity of Scytonema photobionts has never been confirmed by culturing or sequencing. We investigated the phylogenetic placement of presumed Scytonema photobionts and unicellular morphotypes previously assigned to Chroococcus, from tropical Dictyonema, Acantholichen, Coccocarpia, and Stereocaulon lichens. While we confirm that filamentous and unicellular photobiont morphotypes belong to a single clade, this clade does not cluster with Scytonema but represents a novel, previously unrecognized, highly diverse, exclusively lichenized lineage, for which the name Rhizonema is available. The phylogenetic structure observed in this novel lineage suggests absence of coevolution with associated mycobionts at the species or clade level. Instead, highly efficient photobiont strains appear to have evolved through photobiont sharing between unrelated, but ecologically similar, coexisting lineages of lichenized fungi ("lichen guilds"), via the selection of particular photobiont strains through and subsequent horizontal transfer among unrelated mycobionts, a phenomenon not unlike crop domestication.     

Cyanobacterial persistence and influence on microbial community dynamics over 15 years in induced biocrusts

Biocrusts provide numerous ecological functions in drylands. Recovering biocrusts via cyanobacterial inoculation recently gathered interest for ecological restoration, yet it still lacks long-term experiments to unravel biocrust community dynamics. To examine how cyanobacterial inoculants influenced local microbial community and biocrust development, we observed a 2 km² (Qubqi Desert, China) inoculation experiment after 10 and 15 years, following biocrust formation. Our results revealed that biocrust development was in line with ecological regime shift, providing evidence for biocrust community succession, from cyanobacteria- to moss-dominated types. Associated with biocrust development, microbial communities differed significantly with less specialists compared to shifting sands. Cyanobacterial community analysis showed that Microcoleus vaginatus and Scytonema javanicum are an ideal inoculating model, as they were still dominating the community after 15 years since inoculation, while other nitrogen-fixing cyanobacteria occurred profusely with biocrust development. Biocrust community composition combined with thickness, Chl-a and exopolysaccharide measurements revealed the large variation of cyanobacterial ecological functions along biocrust development, suggesting a main function shift: from carbon fixation associated with exopolysaccharide secretion in bare sandy soils to nitrogen fixation in developed biocrusts. This large-scale field study verifies that cyanobacterial inoculation accelerates biocrust development and forwards succession, shaping the biocrust community composition over a long time.     

Biological soil crust (BSC) is an effective biofertilizer on Vigna mungo (L.)

Biological soil crusts (BSC) (Cyanobacteria) play an important role in the soil nitrogen fixation and soil stabilization. However, limited researches were carried out about the diversity and distribution of Bio crust in sacred forests. The study aims to identify the distribution of cyanobacteria in biological soil crusts from different sacred groves of Ariyalur and Pudukottai, Tamil Nadu. We identified following microbes of Microcoleus, Scytonema, Anabaena and Nostoc in biological soil crust. A surface experiment was conducted for the efficacy of biological soil crusts on crops seedling growths. The efficacy was assessed by measuring the root & shoot length, dry & fresh weight of the plant, total chlorophyll, protein & carbohydrate content, with reducing sugar. The plant growth was higher in biocrust inoculated pot than the control. In corresponding nitrogenase activity was determined by the acetylene reduction assay, and phytohormones documentation was executed by the high-performance liquid chromatography. The results indicating that the biological soil crusts are the promising factors influencing the plant growth by plant growth-stimulating auxins and nitrogenase activity.     

Consortial N2 fixation: a strategy for meeting nitrogen requirements of marine and terrestrial cyanobacterial mats

Abstract: Four microbial mat-forming, non-axenic, strains of the non-heterocystous, filamentous, cyanobacterial genus Microcoleus were maintained in culture and examined for the ability to fix atmospheric nitrogen (N₂). Each was tested for nitrogenase activity using the acetylene reduction assay (ARA) and for the presence of the dinitrogenase reductase gene ( nifH ), an essential gene for N₂ fixation, using the polymerase chain reaction (PCR). The Microcoleus spp. cultures were incapable of growth without an exogenous nitrogen source and never exhibited nitrogenase activity. Attempts to amplify a 360-bp segment of the nifH gene using DNA purified from the cyanobacterial cultures did not produce any cyanobacteria-specific nifH sequences. However, several non-cyanobacterial homologous nifH sequences were obtained. Phylogenetic analysis showed these sequences to be most similar to sequences from heterotrophic bacteria isolated from a marine microbial mat in Tomales Bay (California, USA), and bulk DNA extracted from a cryptobiotic soil crust in Moab (Utah, USA). Microcoleus spp. dominated the biomass of both systems. Cyanobacteria-specific 16S rDNA sequences obtained from the cultured cyanobacterial strains demonstrate that the lack of cyanobacteria-specific nifH sequences was not due to inefficiency of extracting Microcoleus DNA. Hence, both the growth and genetic data indicate that, contrary to earlier reports, Microcoleus spp. appear incapable of fixing N₂ because they lack at least one of the requisite genes for this process. Furthermore, our study suggests epiphytic N₂-fixing bacteria form a diazotrophic consortium with these Microcoleus spp. and are likely key sources of fixed N₂ generated within soil crusts and marine microbial mats.     

古尔班通古特沙漠生物结皮微鞘藻分类学研究

实验研究了从古尔班通古特沙漠生物土壤结皮中分离纯化培养出的11株与微鞘藻(Microcoleus)形态接近的丝状蓝藻,通过形态特征、16S rRNA和ITS二级结构相结合的多相分析方法对其进行分类学研究。研究结果表明,实验藻株隶属于微鞘藻科(Microcoleaceae)的微鞘藻属(Microcoleus)和束脉藻属(Symplocastrum),其中包括2个中国新记录种:斯坦微鞘藻(Microcoleus steenstrupii)和细长束脉藻(Symplocastrum flechtnerii),另外还有具鞘微鞘藻(Microcoleus vaginatus)和类似斯坦微鞘藻的存疑物种。藻丝多少与排列方式、细胞大小与末端细胞形状,以及16S rRNA系统发育位置是确定微鞘藻(Microcoleus)与束脉藻(Symplocastrum)属于不同物种的关键依据, ITS二级结构是区分属内不同物种的重要参考。     

Molecular characterization of endolithic cyanobacteria inhabiting exposed dolomite in central Switzerland

The phototrophic microbial community inhabiting exposed dolomite in the alpine Piora Valley (Switzerland) forms a distinct endolithic bilayer that features adjacent red dolomite (exterior) and green dolomite (interior) layers that are c. 0.5-1 mm below the rock surface. Characterization of the community, with an emphasis on cyanobacteria, was conducted with culture-dependent and -independent approaches. Direct microscopy of green dolomite revealed four distinct morphotypes consistent with Chlorophyta genera Chlorella and Stichococcus and the Cyanobacterial genera Nostoc and Calothrix, whereas only Stichococcus and Nostoc were observed in the red dolomite. Enrichment in BG-11 media resulted in the growth of Chlorella and Stichococcus. Denaturing gradient gel electrophoresis (DGGE) analysis of DNA extracted from the enrichment revealed two dominant phylotypes with sequence similarity to Chlorella osrokiniana chloroplast and the cyanobacteria genus Leptolyngbya. 16S rRNA gene-based DGGE analysis of DNA extracted directly from both layers indicated that although both layers harboured phylotypes most similar to the Cyanobacterial genera Nostoc, Chroococcidiopsis, and Microcoleus, and the Chlorophyte Stichococcus bacillaris, the two layers also harboured unique genera such as Scytonema, and Symploca (red, external layer of dolomite) and Chlorella (green, internal layer of dolomite). The unique community structure of each layer suggests a selection process directed by the pressures of the endolithic environment. We conclude that the overall composition of the phototrophic community closely resembles that of endolithic communities located in extreme habitats, suggesting that a cosmopolitan community inhabits this defined niche.     

Nitrogenase activity of blue-green algae on seasonally flooded soils in Bangladesh

Blue-green algal communities formed an extensive cover on soils at five deepwater rice-growing locations in Bangladesh during the month before the arrival of floodwater. The principal taxa wereAnabaena, Cylindrospermum, Lyngbya, Microcoleus, Nostoc, Prophyrosiphon notarisii, Scytonema mirabile andTolypothrix byssoidea. One of two locations studied after the floodwaters had receded also had an extensive cover, mainlyScytonema mirabile. Nitrogenase activity assayed at mid-day was from one to three orders of magnitude higher per unit area of community than bare soil. Nostoc showed higher activity than Tolypothrix, whether expressed per unit area or biomass. Whole field estimates of N₂ fixed by blue-green algal communities during the pre-flood period ranged from 1.0 to 10.2 kg N ha^–1. Much of this is probably not recycled until floodwaters cover the fields. However N₂ fixed after floodwaters have receded is probably recycled rapidly due to ploughing.     

Watering, Fertilization, and Slurry Inoculation Promote Recovery of Biological Crust Function in Degraded Soils

Biological soil crusts are very sensitive to human-induced disturbances and are in a degraded state in many areas throughout their range. Given their importance in the functioning of arid and semiarid ecosystems, restoring these crusts may contribute to the recovery of ecosystem functionality in degraded areas. We conducted a factorial microcosm experiment to evaluate the effects of inoculation type (discrete fragments vs slurry), fertilization (control vs addition of composted sewage sludge), and watering frequency (two vs five times per week) on the cyanobacterial composition, nitrogen fixation, chlorophyll content, and net CO₂ exchange rate of biological soil crusts inoculated on a semiarid degraded soil from SE Spain. Six months after the inoculation, the highest rates of nitrogen fixation and chlorophyll a content were found when the biological crusts were inoculated as slurry, composted sewage sludge was added, and the microcosms were watered five times per week. Net CO₂ exchange rate increased when biological crusts were inoculated as slurry and the microcosms were watered five times per week. Denaturing gradient gel electrophoresis fingerprints and phylogenetic analyses indicated that most of the cyanobacterial species already present in the inoculated crust had the capability to spread and colonize the surface of the surrounding soil. These analyses showed that cyanobacterial communities were less diverse when the microcosms were watered five times per week, and that watering frequency (followed in importance by the addition of composted sewage sludge and inoculation type) was the treatment that most strongly influenced their composition. Our results suggest that the inoculation of biological soil crusts in the form of slurry combined with the addition of composted sewage sludge could be a suitable technique to accelerate the recovery of the composition and functioning of biological soil crusts in drylands.     

Morphological variability in selected heterocystous cyanobacterial strains as a response to varied temperature, light intensity and medium composition

The effect of temperature, light and nutrient composition on morphological traits was determined in seven nostocacean cyanobacteria (Anabaena planctonica, A. sphaerica var. conoidea, A. spiroides, Aphanizomenon gracile, Nostoc sp., Scytonema sp., and Tolypothrix sp.). Their morphological variability was high but only some of the features showed changes reflecting varied growth conditions. The frequency of heterocyst occurrence decreased with increasing nitrogen concentration. Within the range studied, the effect of temperature on heterocyst frequency of Tolypothrix sp. and planktonic Anabaena strains could be fitted by a normal curve with a clear optimum while linear correlation was found in Aphanizomenon gracile. T-and S-type branching was observed in both Scytonema sp. and Tolypothrix sp. strains. T-type branching was found to be markedly dependent on nitrogen concentration. The abundance of necridic cells of Tolypothrix sp. increased linearly with temperature and light intensity. Regularity of trichome coiling of A. spiroides depended on culture medium, suggesting that nutrient composition may be the main controlling factor. In contrast, the effect of the experimental conditions on the dimensions of vegetative cells and heterocysts was weak. Their variability was markedly higher within each experimental treatment than between treatments.     

ROLE OF ULTRAVIOLET RADIATION IN MAINTAINING THE THREE-DIMENSIONAL STRUCTURE OF A CYANOBACTERIAL MAT COMMUNITY AND FACILITATING NITROGEN FIXATION

Cyanobacterial mat communities were collected in the mangrove forest bordering the Grand Cul de Sac Marin, Guadeloupe, French West Indies, which supports a community of nitrogen fixing cyanobacterial mats established on the trunk and branches of black mangrove ( Avicennia germinans L.). This study presents results that are focused on the mat community and the physiological and morphological adaptations to UV radiation. The dominant surface species of the mat, Nostoc cf commune Vaucher and Scytonema sp., possessed the UV-shielding pigment scytonemin. Mats grown on medium D agar without nitrogen under photosynthetically active radiation (PAR) only, rapidly became disorganized compared with those exposed to PAR + UV-A (320– 400 nm) + UV-B (280–320 nm) irradiation. Concurrent with disorganization, acetylene reduction activity (ARA = one third of N₂ reduction) was severely reduced, whereas mats irradiated with PAR + UV-A + UV-B maintained high ARA activity. Mats incubated for 27 days under PAR + UV-A + UV-B then exposed to PAR only exhibited a 68% stimulation of ARA, whereas ARA values were 33% inhibited in mats incubated with PAR only and then exposed to PAR + UV-A + UV-B. This favorable equilibrium was facilitated by the mats' three-dimensional structure in which the most UV-resistant species, N. commune , covers the surface with UV-sensitive species below this protective covering. The UV stressor was essential for the maintenance of mat structure and ARA.     

念珠藻生长特性研究

研究了念珠藻的生长周期、固氮特性、生长条件等生长特性.得出本实验念珠藻可以固氮;磷能有效制约其生长;需要长时间的光照等结果.     

Associative cyanobacteria isolated from the roots of epiphytic orchids

Associative cyanobacteria were isolated from the rhizoplane and velamen of the aerial roots of the epiphytic orchids Acampe papillosa, Phalaenopsis amabilis, and Dendrobium moschatum and from the substrate roots of Acampe papillosa and Dendrobium moschatum. Cyanobacteria were isolated on complete and nitrogen-free variants of BG-11 medium. On all media and in all samples, cyanobacteria of the genus Nostoc predominated. Nostoc, Anabaena, and Calothrix were isolated from the surface of the A. papillosa aerial roots, whereas the isolates from the substrate roots were Nostoc, Oscillatoria, and representatives of the LPP-group (Lyngbia, Phormidium, and Plectonema, incapable of nitrogen fixation). On the D. moschatum substrate roots, Nostoc and LPP-group representatives were also found, as well as Fischerella. On the aerial roots of P. amabilis and D. phalaenopsis grown in a greenhouse simulating the climate of moist tropical forest, cyanobacteria were represented by Nostoc, LPP-group, and Scytonema in the D. phalaenopsis and by Nostoc, Scytonema, Calothrix, Spirulina, Oscillatoria, and the LPP-group in P. amabilis. For D. moschatum, the spectra of cyanobacteria populating the substrate root zhizophane and the substrate (pine bark) were compared. In the parenchyma of the aerial roots of P. amabilis, fungal hyphae and/or their half-degraded remains were detected, which testifies to the presence of mycorrhizal fungi this plant. This phenomenon is attributed to the presence of a sheath formed by cyanobacteria and serving as a substrate for fungi.     

几种固沙荒漠藻的温度适应性

蓝藻结皮在干旱和半干旱区广泛分布,它们在环境状态的维持和改良过程中发挥着重要的作用。在荒漠藻固沙技术的应用过程中,温度不仅影响藻类在培养池中的培养,还影响接种后藻类的生长。因此,摸清优势固沙藻类的温度生长特征及对不同温度的生长适应性,对于接种蓝藻固沙技术的应用具有积极的意义。实验分3部分:研究了室温培养条件下3种荒漠优势蓝藻-具鞘微鞘藻(Microcoleus vaginatus)、爪哇伪枝藻(Scytonema javanicum)、纤细席藻(Phormidium tenue)的生长曲线,在不同温度(2、5、10、15、25、35℃)、开放式载体培养状态下3种蓝藻的生长状况及形态观察,以及爪哇伪枝藻在不同温度(10、15、20、25、30℃)培养条件下的光合活性、光合色素含量和伪枝藻素含量的变化。实验结果表明:(1)在液体培养基中,纤细席藻生长速率最快,高于具鞘微鞘藻和爪哇伪枝藻;(2)开放式载体培养条件下,藻株的生长速率低于液体培养,因此荒漠优势藻类的培养优先选择液体培养,具鞘微鞘藻和爪哇伪枝藻不易被细菌污染,纤细席藻容易受到细菌污染,在培养该藻株时要考虑添加抗生素类药物抑制细菌过度繁殖;(3)爪哇伪枝藻短期培养(18d)宜选择相对较高的培养温度(25-30℃),而长期培养(30d)宜选择相对较低的培养温度(15-20℃)。