Evidence for NH4+ switch-off regulation of nitrogenase activity by bacteria in salt marsh sediments and roots of the grass Spartina alterniflora

The regulatory effect of NH4+ on nitrogen fixation in a Spartina alterniflora salt marsh was examined. Acetylene reduction activity (ARA) measured in situ was only partially inhibited by NH4+ in both the light and dark after 2 h. In vitro analysis of bulk sediment divided into sediment particles, live and dead roots, and rhizomes showed that microbes associated with sediment and dead roots have a great potential for anaerobic C2H2 reduction, but only if amended with a carbon source such as mannose. Only live roots had significant rates of ARA without an added carbon source. In sediment, N2-fixing mannose enrichment cultures could be distinguished from those enriched by lactate in that only the latter were rapidly inhibited by NH4+. Ammonia also inhibited ARA in dead and live roots and in surface-sterilized roots. The rate of this inhibition appeared to be too rapid to be attributed to the repression and subsequent dilution of nitrogenase. The kinetic characteristics of this inhibition and its prevention in root-associated microbes by methionine sulfoximine are consistent with the NH4+ switch-off-switch-on mechanism of nitrogenase regulation.     

Lacunal allocation and gas transport capacity in the salt marsh grass Spartina alterniflora

Summary Lacunal allocation as the fraction of the total cross sectional area of leaves, stem bases, rhizomes, and roots was determined in both tall and short growth forms of Spartina alterniflora collected from natural monospecific stands. The results indicate that in both growth forms lacunal allocation is greater in stem bases and rhizomes than in leaves and roots and that tall form plants allocate more of their stem and rhizome to lacunae than short form plants.Measurements made in natural stands of Spartina alterniflora suggest that total lacunal area of the stem base increases with increasing stem diameter and that stem diameter increases with increasing plant height and above-ground biomass. However, the fraction of cross section allocated to lacunae was relatively constant and increased only with the formation of a central lacuna.Experimental manipulations of surface and subsurface water exchange were carried out to test the influence of flooding regime on aerenchyma formation. No significant differences in lacunal allocation were detected between plants grown in flooded (reduced) and drained (oxidized) sediments in either laboratory or field experiments. While aerenchyma formation in Spartina alterniflora may be an adaptation to soil waterlogging/anoxia, our results suggest that lacunal formation is maximized as a normal part of development with allocation constrained structurally by the size of plants in highly organic New England and Mid-Atlantic marshes.The cross sectional area of aerenchyma for gas transport was found to be related to the growth of Spartina alterniflora with stands of short form Spartina alterniflora exhibiting a lower specific gas transport capacity (lacunal area per unit below ground biomass) than tall form plants despite having a similar below-ground biomass supported by a 10 fold higher culm density. The increased specific gas transport capacity in tall vs. short plants may provide a new mechanism to explain the better aeration, higher nutrient uptake rates and lower frequency of anaerobic respiration in roots of tall vs. short Spartina alterniflora.     

Oxygen loss from Spartina alterniflora and its relationship to salt marsh oxygen balance

Spartina alterniflora has been reported to lose significant amounts of oxygen to its rhizosphere with potentially important effects on salt-marsh biogeochemical cycling and plant productivity. The potential significance of this oxidative pathway was evaluated using laboratory split-chamber experiments to quantify oxygen loss from intact root systems under a wide variety of pre-treatment and incubation conditions including antibiotics to inhibit microbial respiration. The aerenchyma system of S. alterniflora was found to transport O₂, N₂, Ar, and CH₄ from above-ground sources to its below-ground roots and rhizomes. While non-respiratory gases were observed to move from the lacunae to water bathing the root systems, net O₂ loss did not occur; instead oxygen present outside of the roots/rhizomes was consumed. Net oxygen loss was found when resistance to gas transport was reduced in the lacunae-rhizosphere pathway by placing the root systems in a gas phase and when plant respiration was significantly reduced. Root system respiration appeared to be the major variable in the plant oxygen balance. When root and rhizome respiration was inhibited using poisons or lowered by cooling, the oxygen deficit was greatly reduced and oxygen loss was indicated. The effect of seasonal temperature changes on root system oxygen deficit presents a possible explanation as to why Spartina produces root systems with respiration rates that cannot be supported by gas transport. Overall, while oxygen loss from individual plant roots is likely, integrating measured root system oxygen loss with geochemical data indicates that the mass amount of oxygen lost from S. alterniflora root systems is small compared to the total oxygen balance of vegetated salt marsh sediments.     

Estimation of primary production using five different methods in a Spartina alterniflora salt marsh

The aboveground production of Spartina alterniflora in a salt marsh in Barataria Bay, Louisiana, USA was estimated using five different harvest methods: peak standing crop (PSC), Milner-Hughes, Smalley, Wiegert-Evans, and Lomnicki et al., and a non-destructive method based on measurement of stem density and longevity. Annual production estimates were 831 ± 41, 831 ± 62, 1231 ± 252, 1873 ± 147 and 1437 ± 96 g dry wt m^–2 for each method, respectively. The average longevity of individually tagged young shoots was 5.2 ± 0.2 months, equivalent to an annual turnover rate of 2.3 crops per year. Among the five methods, Wiegert-Evans and Lomnicki et al. were considered more accurate than the other three because they corrected for mortality losses between sampling times. The Lomnicki et al. method was preferred over the Wiegert-Evans method because of its greater simplicity.     

Analysis of nitrogen-fixing members of the epsilon subclass of Proteobacteria in salt marsh sediments

Based on phylogenetic analysis of clones retrieved from two nifH gene clone libraries that were created using cDNA from suboxic sediment samples obtained from areas densely vegetated with the high-salt marsh plant Spartina patens, a primer set was designed to target nitrogen-fixing bacteria with sequence similarities to members of the epsilon subclass of Proteobacteria. Nested PCR, denaturing gel electrophoresis, and subsequent sequence analysis of reamplified fragments confirmed the specificity of the primer set by retrieving nifH sequences of only putative members of the epsilon subclass of Proteobacteria, all of which were characterized by a highly divergent 27- or 36-bp insertion in both DNA and cDNA.     

Designing an effective clipping regime for controlling the invasive plant Spartina alterniflora in an estuarine salt marsh

Many invasive plants are highly tolerant of disturbances and can invade various habitats, leading to difficulties in managing them. Most of control techniques need to be performed repeatedly, and the efficiency of the same control method may vary along environmental gradients. To develop a control strategy for an invasive plant in heterogeneous environments, it is important not only to optimize the control regime, but also to consider the efficiency in different habitats. We conducted a field experiment to test how clipping regime (i.e., initial treatment timing and treatment interval) and habitat type (i.e., high and low tidal zones) influence the control efficiency for invasive Spartina alterniflora in an estuarine salt marsh. Among the 12 treatments, double-clipping treatment performed in early July and mid September (9-week-interval) had the highest efficiency, which not only eliminated sexual reproduction of S. alterniflora, but also inhibited its vegetative growth in the current and following years. The clipping efficiency in low tidal zone was higher than that in high tidal zone. Therefore, an optimized control strategy for the invasive plants could be established based on the efficient control regime and habitat types. According to our study, a recommended double-clipping control strategy for S. alterniflora is that the initial clipping treatment is performed in the flowering period and the second one is performed 9 weeks later. In order to effectively reduce proliferation of the invasive plant, we suggest that the clipping regime should be applied for at least 2–3 years in low tidal zone and 3–4 years in high tidal zone.     

Effects of salt marsh invasion by Spartina alterniflora on sulfate-reducing bacteria in the Yangtze River estuary,China

To investigate how plant invasion affects sulfate-reducing bacteria (SRB) responsible for sulfate reduction, we conducted a comparative study of diversity and composition of SRB in rhizosphere soils of invasive exotic species (Spartina alterniflora) and two native species (Phragmites australis and Scirpus mariqueter) on Jiuduansha Island located in the Yangtze River estuary, China. Throughout the growing season, profiles of DGGE fingerprints of SRB had distinct variations in relation to phenological stages of these three plant species. The higher richness and abundance of SRB in the rhizospheres of native plants mainly occurred when the plants were in vegetative growth and reproductive stages. However, the higher richness and abundance of SRB also occurred in the late growing season (senescent stage) of S. alterniflora rhizosphere, during which Desulfobulbus, Desulfuromonas, Desulfovibrio, and Firmicutes were dominant. Our results adding to our previous studies suggested that abundant SRB in late stage might have close relationships with decomposition of soil organic matters produced by S. alterniflora.     

Crenarchaeal heterotrophy in salt marsh sediments

Mesophilic Crenarchaeota (also known as Thaumarchaeota) are ubiquitous and abundant in marine habitats. However, very little is known about their metabolic function in situ. In this study, salt marsh sediments from New Jersey were screened via stable isotope probing (SIP) for heterotrophy by amending with a single ¹³C-labeled compound (acetate, glycine or urea) or a complex ¹³C-biopolymer (lipids, proteins or growth medium (ISOGRO)). SIP incubations were done at two substrate concentrations (30–150 μM; 2–10 mg ml⁻¹), and ¹³C-labeled DNA was analyzed by terminal restriction fragment length polymorphism (TRFLP) analysis of 16S rRNA genes. To test for autotrophy, an amendment with ¹³C-bicarbonate was also performed. Our SIP analyses indicate salt marsh crenarchaea are heterotrophic, double within 2–3 days and often compete with heterotrophic bacteria for the same organic substrates. A clone library of ¹³C-amplicons was screened to find matches to the ¹³C-TRFLP peaks, with seven members of the Miscellaneous Crenarchaeal Group and seven members from the Marine Group 1.a Crenarchaeota being discerned. Some of these crenarchaea displayed a preference for particular carbon sources, whereas others incorporated nearly every ¹³C-substrate provided. The data suggest salt marshes may be an excellent model system for studying crenarchaeal metabolic capabilities and can provide information on the competition between crenarchaea and other microbial groups to improve our understanding of microbial ecology.     

江苏沿海互花米草(Spartina alterniflora Loisel)盐沼扩展过程的遥感分析

互花米草(SpartinaalternifloraLoisel)自1982年在江苏沿海栽种以来,在江苏沿海已形成了大片互花米草盐沼。本文通过对历年TM卫星相片上互花米草盐沼的识别、判读及统计,认为江苏沿海互花米草盐沼的年扩展速度在早期较慢,仅为23 4%,中期较快为89%,后期又减慢为48%。结合野外调查,绘制出了江苏沿海互花米草的分布图。此外,就互花米草盐沼对江苏沿海湿地植被演替的影响进行了讨论。     

Effects of drainage and soil organic content on growth of Spartina alterniflora (Poaceae) in an artificial salt marsh mesocosm

An artificial salt marsh mesocosm was constructed using 680-L polypropylene tanks to determine the effect of soil drainage depth and organic content on growth and rhizome proliferation of the salt marsh smooth cordgrass Spartina alterniflora. Soil drainage depth had no effect on accumulation of aerial or subsurface plant tissue, but tanks that had 2.5% soil organic content supported enhanced aerial tissue and rhizome growth compared to tanks that contained sand alone. We propose a mathematical model for predicting the mass of photosynthetically significant leaf tissue without cutting and drying leaves. Implications of these findings for salt marsh creation projects are discussed.     

互花米草入侵对沿海湿地甲烷排放的影响

采集互花米草不同入侵年限(8、11和15年)的原状土壤,采用盆栽试验,研究了土壤有机碳含量对沿海湿地CH4排放的影响。结果表明,土壤有机碳含量随着互花米草入侵年限的增加而增加。在植物生长季,互花米草入侵15年的土壤有机碳含量为12.97g·kg-1,土壤CH4排放通量为2.94mg·m-2·h-1,显著高于入侵年限为8和11a(有机碳含量为8.11和9·16g·kg-1)的土壤,其土壤CH4排放通量分别为1.95和2.34mg·m-2·h-1。这主要是由于随着土壤有机碳含量提高,不仅为产甲烷菌提供了更多底物,同时也促进了产甲烷菌数量增加,从而导致更多CH4排放。因此,在评价互花米草入侵的综合环境效应时,需要兼顾土壤固碳能力和温室气体排放。     

Seasonal patterns of daily net photosynthesis,transpiration and net primary productivity of Juncus roemerianus and Spartina alterniflora in a Georgia salt marsh

Summary Studies of the seasonal CO₂ and water vapor exchange patterns of Juncus roemerianus and Spartina alterniflora were conducted in an undisturbed marsh community on Sapelo Island, Georgia. Daily patterns of net photosynthesis, transpiration, leaf diffusive conductance and water-use efficiency in response to ambient conditions were monitored on intact, in situ plants. Net primary productivity was calculated from the daytime CO₂ fixation totals, nighttime CO₂ loss, leaf standing stock and aboveground to belowground biomass ratios for each plant type.The tall form of S. alterniflora had higher rates of photosynthesis and higher water-use efficiency values which, in conjunction with low respiratory losses and large leaf standing crop, results in high values of net primary productivity. The environmental factors in the marsh which permit these physiological responses occur in less than 10% of the marsh. Overall, the physiological capabilities of the short form of S. alterniflora were reduced in comparison to the tall form, but the combination of environmental factors which determine the physiological responses of this form occur in a much greater portion of the marsh, and the short form of S. alterniflora dominates the Sapelo Island marshes.The response patterns of the C₃ species, J. roemerianus, differed somewhat from the height forms of S. alterniflora. A large, seasonally constant leaf standing crop coupled with moderate rates of photosynthesis resulted in a net primary productivity value which was between the tall and short height forms of S. alterniflora. However, as with the tall S. alterniflora, the environmental conditions under which this high productivity and high water loss rate can be sustained are restricted to specific regions of the environmental gradient in the marsh.Contribution No. 435 from the University of Georgia Marine Institute     

In situ biogenic silica variations in the invasive salt marsh plant, Spartina alterniflora: A possible link with environmental stress

Aims

Higher plants are an understudied component of the global silicon cycle; they absorb silicic acid (dSi) which is stored as biogenic silica (bSiO₂). Si is believed to alleviate physical, chemical, and biological stresses such as storms, high salinity, heavy metal toxicity, grazing, and disease. We investigated a Si-accumulating invasive species growing in the tidal marshes of the Bay of Brest (France), viz., Spartina alterniflora. Our objectives were to determine (1) where and when bSiO₂ accumulates in the plant during its life cycle, (2) whether this accumulation varies with abiotic factors: wave action, estuarine salinity, and duration of immersion, and (3) if the accumulation was limited by dSi availability in marsh porewater.

Methods

A 2?years field survey permitted to sample plants which were analyzed for there bSiO₂ concentrations. Sediment cores were sampled seasonally and the dSi concentrations in the porewater were measured from 0 to 10?cm.

Results

bSiO₂ accumulated more in mature leaves than in other organs. There was a strong linear relationship between bSiO₂ concentration and plant length. bSiO₂ concentrations did not increase, but rather decreased as a function of exposure to the three abiotic factors tested. dSi availability was not significantly different for each of the tested sites and dSi profiles did not exhibit huge losses in the root zone.

Conclusions

Our evidence suggests that dSi availability did not seem to be a limiting factor. bSiO₂ did not increase with increasing abiotic stresses but was strongly correlated with growth. Hence, S. alterniflora is likely to have other adaptive strategies for dealing with environmental stressors but it did not exclude the possible role of Si in alleviating these stresses. If this is the case, there remain intriguing questions about Si uptake, its availability, and its role in silicification and growth.     

Accumulation of proline and glycinebetaine in Spartina alterniflora Loisel. in response to NaCl and nitrogen in the marsh

Summary The possible interaction of high soil salinity and low soil nitrogen content in affecting the growth of Spartina alterniflora Loisel in the high and low marshes of the Eastern U.S. was explored. Throughout the whole growing season, the short plants growing in the high marsh, where there was a higher soil salinity and lower available soil nitrogen, contained more proline and glycinebetaine and showed a lower leaf water potential than the tall plants growing in the low marsh. In both short and tall plants, the growing season, with the highest content occurring in spring and fall. In contrast, the glycinebetaine content in both short and tall plants remained fairly constant throughout the growing season, and was consistently at least 10 fold higher than the proline content. It is estimated that 19–30% of the total leaf nitrogen was in the form of proline and glycinebetaine in the short plants, and 14–27% in the tall plants. Ammonium nitrate fertilization in the field resulted in increased growth, higher proline and glycinebetaine contents, and lower water potentials in the short plants, but had little effect on these parameters in the tall plants. We suggest that in the low marsh, the plants can obtain sufficient nitrogen for osmoregulation and other metabolism. In the high marsh with higher soil salinity and lower nitrogen content, the plants have to allocate a even greater proportion of the already limited nitrogen supply for osmoregulation. Thus, nitrogen available for osmoregulation and other nitrogen-requiring metabolism is insufficient, resulting in reduced growth.     

Tiller dynamics of Spartina maritima in successional and non-successional mediterranean salt marsh

Tiller demography was compared in two populations of Spartina maritima present at similar elevations in the coastal saltmarshes of Odiel (Huelva, S.W. Spain). The successional population consisted of colonizing tussocks in a littoral lagoon, and the non-successional population comprised a stable sward that had fringed a major channel for 40 years. At both sites S. maritima was replaced by Arthrocnemum perenne at higher elevation, where sediments were less reducing. Rapid, consistent sediment accretion confirmed the successional nature of the lagoon site but there was little net accretion in the stable sward.Census of permanent quadrats at the successional site chronicled moving concentric waves of high tiller density as tussocks expanded. Initially high densities declined after one year to low values at the end of the second year but they had almost recovered after 3 years. The decline represented a combination of reduced numbers of births and increased numbers of deaths. Tiller densities were substantially higher in the stable sward and showed relatively small fluctuations with time. The underlying risk of tiller mortality was similar in the two populations for much of the time but after two years there was increased mortality, mainly associated with flowering, at the successional site; very few tillers flowered in the sward. This mortality contributed to a shift to a younger age structure in the successional population.Data aggregated over consecutive 3-monthly periods were examined for density dependence. None was found in the successional population. In the sward population there was evidence of density-dependent adult and juvenile mortality of tillers, particularly over the first 18 months of the study, when there were compensatory responses to subtle variations in density. The lack of density dependence and relatively low peak density of about 2000 m^-2 near to the leading edges of the expanding tussocks at the successional site suggest that tiller placement there was regulated mainly by physiological mechanisms affecting rhizome growth and bud development in well integrated clones.     

Microbial carbon monoxide consumption in salt marsh sediments

We have examined sediments from a fringing salt marsh in Maine to further understand marine CO metabolism, about which relatively little is known. Intact cores from the marsh emitted CO during dark oxic incubations, but emission rates were significantly higher during anoxic incubations, which provided evidence for simultaneous production and aerobic consumption in surface sediments. CO emission rates were also elevated when cores were exposed to light, which indicated that photochemical reactions play a role in CO production. A kinetic analysis of marsh surface sediments yielded an apparent K(m) of about 82 ppm, which exceeded values reported for well-aerated soils that consume atmospheric CO (65nM). Surface (0-0.2 cm depth interval) sediment slurries incubated under oxic conditions rapidly consumed CO, and methyl fluoride did not inhibit uptake, which indicated that neither ammonia nor methane oxidizers contributed to the observed activity. In contrast, aerobic CO uptake was inhibited by additions of readily available organic substrates (pyruvate, glucose and glycine), but not by cellulose. CO was also consumed by surface and sub-surface sediment slurries incubated under anaerobic conditions, but rates were less than during aerobic incubations. Molybdate and nitrate or nitrite, but not 2-bromoethanesulfonic acid, partially inhibited anaerobic uptake. These results suggest that sulfidogens and acetogens, but not dissimilatory nitrate reducers or methanogens, actively consume CO. Sediment-free plant roots also oxidized CO aerobically; rates for Spartina patens and Limonium carolinianum roots were significantly higher than rates for Spartina alterniflora roots. Thus plants may also impact CO cycling in estuarine environments.